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Business Intelligence: Building Your First Cube. I like to science essay, introduce people to Microsoft SQL Server Analysis Services by creating a simple cube that contains a sampling of Close and Fictional Narratives the Dominant Narrative their own data. I have to be careful when I do this, though, because I have been pushed out of my chair on more than one occasion when new users got excited about what they were seeing and science essay wanted to dive right in to explore the data in ways they never before thought possible. Whether you have data of online your own you want to explore more easily or a data-hungry user community to support, you'll find SQL Server Analysis Services (SSAS) provides the tools. This article is the third installment of science essay a series; the first two articles appeared in the August 2009 issue and are available at This series will help you develop a better understanding of grendel book business intelligence (BI) by using the science essay, components available in Microsoft SQL Server 2008 as you build a simple BI solution. Capital Pricing! Because I'll be referring to concepts and science terminology introduced in the previous two articles, I recommend you take the time to enslaved, review those articles first if you haven't read them already. In Planning Your First BI Solution, I explained the essay, advantages of building a data mart compared with retrieving data directly from grendel book, a corporate database. I also provided instructions for downloading the sample databases you'll need in order to science essay, follow the steps described in these articles. Grendel Book! Then Derek Comingore, in Building a Data Foundation for science, a BI Solution, explained how to cola vs pepsi sales, use Integration Services to essay, populate a data mart with data from your corporate database. Mondays Kingston Jamaica Essay! In this article, I'll show you how to build a cube based on a similar data mart design.

My first article summarized the reasons to science, consider moving data out of your corporate database and into a structure called a data mart. Pricing! While querying a data mart certainly has its benefits, and is probably sufficient for essay, some scenarios, there are additional benefits to capital pricing model, be realized by repackaging that data one more time into a structure called a cube. Science! In fact, the data mart exists as an intermediate stop as the data makes its way to its ultimate destination in Mondays Kingston Essay, the cube, so it's still important in the overall solution. Essay! An added benefit of building a cube to house your data is coca cola that you can centralize the business rules for calculations that you can't easily store in a relational data mart. Essay! Furthermore, the pricing, structure of the cube makes it much easier to essay, write queries to thinking, compare data year over year, or to create cumulative values such as year-to-date sales. Moreover, you gain the ability to transparently manage aggregated data in the cube. To improve query performance in a relational data mart that contains a lot of data, database administrators often create summary tables to prepare data for queries that don't require transaction-level detail. SSAS creates the science essay, logical equivalent of summary tables (called aggregations) and grendel book keeps them up-to-date. Science Essay! In this article, I'll continue the scenarios introduced in the first article. These scenarios define the basic analysis objectives for the BI solution you're building throughout this series. The solution must be able to show which sales channel is more profitable for Adventure Works, Internet Sales or Reseller Sales, and whether sales trends indicate that demand is growing or declining for certain products.

Your source data for enslaved africans, the BI solutions is the AdventureWorksDW2008 database, which applies the dimensional modeling and ETL principles described in essay, Derek's article. You can download the sample databases used to build the solution for Adventure Works from CodePlex at capital asset model With a data mart available as a source, you're ready to science essay, build an tu clothing online, Analysis Services 2008 database. Preparing Your Analysis Services Database. Science! Your first step is to create a project in lord flies, Business Intelligence Development Studio (BIDS). Essay! Launch BIDS from the Microsoft SQL Server 2008 program group, click File | New | Project. In the grendel book, New Project dialog box, select Analysis Services Project. In the Name text box, type ssas_TECHNET_AW2008 and, if you like, change the location for your project.

Click OK to create the project. Now add a data source to define the science essay, connection string for your data mart. Lord Flies Beast! In Solution Explorer, right-click the Data Sources folder and science click New Data Source. Coca Cola! In the science essay, Data Source Wizard, click Next on grendel book the Welcome page if it hasn't been disabled. On the Select How to essay, Define the capital asset, Connection page, click New to science, set up a new connection. In the tu clothing, Connection Manager, the essay, default is the SQL Server Native Client, which is thinking education correct for this project, though you could also use an science essay, OLE DB or .NET provider to access other types of data. In Jamaica! To define the connection, type the name of your server in essay, the Server Name text box or select it from the tu clothing, drop-down list, then select AdventureWorksDW2008 in the database drop-down list and science essay click OK. When you're back in grendel book, the Data Source Wizard, click Next. On the Impersonation Information page, select Use the service account option. The service account is used to essay, read data from the source when loading data into your SSAS database and must have read permissions to grendel book, do so.

Click Next and then Finish to essay, complete the tu clothing online, wizard. Science Essay! Next, you create a data source view (DSV) as an education, abstraction of the science essay, tables (or views) from the lord of the flies, data source you want to science, use to define dimensions and cubes. You can make changes to of Historical and Fictional Narratives Theories: Challenging, the DSV without modifying the underlying data source, which is science handy if you have only read permissions to the data mart and can't make changes at the source. In Solution Explorer, right-click the Data Source Views folder and then click New Data Source View. Click Next on critical thinking education the Welcome page if necessary. Science Essay! On the Select a data source page, select the data source you just added to thinking education, the project and click Next. Add objects to science essay, the DSV by double-clicking each table or view you need. To create a DSV that helps you answer the BI questions posed at the beginning of this article, add the thinking education, following tables to science essay, the DSV: DimDate, DimProduct, DimProductCategory, DimProductSubcategory, FactInternetSales and FactResellerSales. Enslaved! While you're learning how to work with SSAS, it's a good idea to start with a simple DSV like this one.

You can always add more tables later if you need to support other questions with your BI solution. Click Next in the Data Source View Wizard when you've finished adding tables and then click Finish. I recommend you simplify the names of your tables by selecting each one in the Tables pane of the DSV designer and removing the science, Dim and Fact prefixes from the FriendlyName property of coca cola vs pepsi each table. The wizards you use to create dimensions and cubes will then use the value in the FriendlyName property to assign names to essay, objects. Thinking Education! The complete DSV is science shown in Figure 1 . In addition to capital model, changing a table's properties in the DSV, you can define logical primary keys or logical relationships between tables if the physical source does not have them defined already. SSAS won't be able to science essay, display data correctly without these definitions in place physically in the data layer or logically in the DSV.

You can also add a named calculation, which is grendel book like adding a derived column to a view, or replace the science essay, table object with a named query, which is grendel book similar to science, creating a view. Readings And Fictional Narratives Of Conspiracy! In preparation for building the date dimension in SSAS, you need to essay, add two named calculations to the Date table -- Quarter and Month -- to concatenate the year column with the quarter and month columns. Otherwise, data will not roll up correctly by month, quarter and year when you want to asset, view summarized data. To add the named calculation for essay, Quarter, right-click on the Date table in coca, either the science, designer or the Tables pane and enslaved click New Named Calculation. In the Column Name text box, type Quarter. Essay! In the thinking, Expression text box, type the following expression: 'Qtr ' + convert(char(1), CalendarQuarter) + ' ' + convert(char(4), CalendarYear) Click OK and science now repeat these steps to add the named calculation for Close Readings of Historical and Fictional Narratives Challenging, Month, using the essay, following expression: left(EnglishMonthName, 3) + ' ' + convert(char(4), CalendarYear) Notice that the critical, Date table in the designer (shown in Figure 2 ) displays your named calculations with a calculator icon to essay, differentiate them from columns physically in the table. Tu Clothing! You can review the effect of adding the essay, named calculations by right-clicking the table and grendel book clicking Explore Data.

A new window opens in BIDS to display the data from the source table. Science! When you scroll all the Uptown Kingston, way to the right, you can see the values for the named calculations. You should always use the Explore Data command to confirm that the named calculations display the science essay, way you intend before you proceed to education, building your dimensions. Science Essay! Defining Dimensions in capital asset model, Analysis Services. Science Essay! After adding dimension tables to the DSV, you're ready to create dimensions in Close of Conspiracy Challenging the Dominant, the SSAS database. As a reminder, you use a dimension to store information about business entities, such as people, places and things.

To be able to answer the science essay, BI questions posed at the beginning of this article, you need to create dimensions for vs pepsi sales, Date and Product. Essay! To add a dimension to online, the project, launch the Dimension Wizard by science essay, right-clicking the Dimensions folder in Solution Explorer and critical selecting New Dimension. On the Select Creation Method page, keep the default selection to essay, use an existing table because your DSV includes the asset model, DimDate table. For those situations when you might build a simple database based on science a small transactional table that includes dates, you might not build out africans a data mart and can instead use one of the other options on this page to science, generate a time table in the data source or on the server. Figure 2 Date Table with Named Calculations.

In either case, you supply a beginning date and an end date for critical thinking, the data in the transactional table. SSAS can create and science populate a physical table in of the beast, the data source it will use to science, populate the Close of Historical and Fictional Narratives of Conspiracy Challenging the Dominant Narrative, dimension maintained in the SSAS database, or it can simply maintain the science essay, dimension exclusively in the SSAS database. Of The Flies! You might choose to generate the physical table when you need to be able to support relational queries to the data source. Click Next to essay, continue. Tu Clothing! On the Specify Source Information page, select Date in the Main Table dropdown list.

Note the Key Column is DateKey. The key column must uniquely identify each record in the table so that SSAS can correctly aggregate or group data when returning query results. Essay! Then, in tu clothing, the Name Column dropdown list, select FullDateAlternateKey. SSAS uses the name column as a label to display to the user instead of displaying the key column. On the Select Dimension Attributes page, you choose attributes to include in the dimension, which you use to define hierarchies or additional grouping labels. Science Essay! Whether you include all attributes from the dimension table depends on Close of Historical and Fictional Narratives Narrative the types of questions you want to support in science essay, your BI solution. Enslaved Africans! Add only as many as you need to science, ensure SSAS performs optimally, to Uptown in Kingston Jamaica Essay, avoid consuming storage space unnecessarily and to avoid confusing users with too many options. For this dimension, select Calendar Year, Quarter and Month. Click Next and then click Finish.

To ensure that each attribute can be uniquely identified, update the science, KeyColumn property. By setting this property to the key columns, you ensure that Month sorts correctly in tu clothing online, numerical sequence rather than alphabetically by month name, which is the science essay, default sort order. To update the KeyColumn property, select Month in the Attributes pane on the left. In the Properties window, click the tu clothing online, KeyColumns property and then the science, ellipsis button. In the Key Columns dialog box, click the left arrow to clear the current assignment and then double-click CalendarYear and MonthNumberOfYear. Now assign a label to tu clothing, this attribute by essay, clicking the Close Narratives Challenging, ellipsis button in the NameColumns property box. Select EnglishMonthName, and science click OK. Repeat these steps to set the KeyColumns property of the Quarter attribute to enslaved, CalendarYear and CalendarQuarter and to set the NameColumns property to science essay, Quarter. The dimension now appears in asset, Solution Explorer as Date.dim and essay the dimension designer (shown in tu clothing online, Figure 3 ) displays in the main workspace area in BIDS. Figure 3 Date Dimension Shown in science essay, the Designer. You can see the dimension with its associated attributes, including the enslaved africans, key attribute, in the Attributes pane on science essay the left.

If you decide later to cola vs pepsi, add more attributes, you can drag them one by one from the Data Source View pane to the Attributes pane. You use the Dimension Wizard only when starting a new dimension, but can make any necessary changes later in the dimension designer. Now use the science essay, Dimension Wizard to cola sales, create the science essay, Product dimension, using Product as the africans, main table and science EnglishProductName as the name column. Lord Flies Beast! Because the science essay, Product dimension is a snowflake schema, the wizard includes an in Essay, additional page to science, confirm that you want to include the tu clothing online, related tables, ProductSubcategory and ProductCategory. Next, add the essay, Color and tu clothing online Size attributes to the dimension. Science Essay! Notice that the tu clothing online, wizard automatically selects the key columns for essay, the snowflake tables, Product Subcategory Key and Product Category Key. You need to tu clothing, add in the corresponding name columns for these attributes, but must first finish the wizard so you can modify the attribute properties.

When the essay, dimension designer displays, select Product Category Key in the Attributes pane. In the Properties, locate the Name property and change the Close of Historical Narratives of Conspiracy the Dominant, name to Category. Essay! Scroll down in sales, the Properties window to essay, locate the NameColumn property. Click in the property text box to display the ellipsis button, click the button, select EnglishProductCategoryName, and click OK. Now repeat these steps to rename Product Subcategory Key to Subcategory and to specify EnglishProductSubcategoryName as its name column. Last, rename the Product Key attribute as Product. Notice the blue wavy line that appears under the dimension name, Product, in the Attributes pane. When you hover your cursor over this line, a warning message appears, Create hierarchies in tu clothing, non-parent child dimensions.

This message is an science essay, example of the best practices warnings built into enslaved africans SSAS 2008 to help you build an science, SSAS database correctly. Return to lord flies, the Date dimension designer, either by clicking the applicable tab in science, the document workspace or by enslaved, double-clicking Date.dim in science, Solution Explorer, and you'll see the lord of the, same warning there. Essay! Adding a hierarchy to grendel book, a dimension is science essay considered a best practice for capital pricing model, several reasons, in essay, particular for flies, user-friendliness and optimization purposes. More specifically, a hierarchy provides a navigation path for users to science, follow from flies, summarized to science essay, detailed data. Cola Sales! Moreover, it can improve query performance by enabling SSAS to science essay, compute and store aggregations in advance of grendel book user queries. For example, if the essay, user wants to online, see reseller sales by science essay, year and grendel book a hierarchy in the Date dimension defines the rollup path from the key attribute (Date Key) to the Calendar Year attribute, SSAS can calculate the sales data by essay, year during processing and then put the Close Readings Narratives of Conspiracy Theories: Challenging the Dominant Narrative, results into permanent storage. This storage of aggregations avoids the science essay, need to thinking, compute each year's sales at science essay query time and is the key difference between retrieving data from critical thinking education, a relational data mart or from science, a multidimensional database like SSAS. To add a hierarchy to the Date dimension, drag the Calendar Year attribute from the asset pricing, Attributes pane to the Hierarchy pane in the dimension designer.

After you add the science, first attribute, a hierarchy object appears with a new empty level below the pricing, attribute you just added. Add the Quarter and Month attributes to the hierarchy by dropping each one into science essay the space for the empty level. Thinking! Next, rename the science essay, hierarchy by right-clicking Hierarchy, then clicking Rename and typing Year. Although you've added a hierarchy, a warning still appears for capital asset, the Date dimension. Hover your cursor over the line to see a new warning, Avoid visible attribute hierarchies for attributes used as levels in user-defined hierarchies. This message suggests you prevent users from viewing an essay, attribute by itself when that attribute has been included in a hierarchy.

In other words, a user should only be able view the lord of the beast, attribute when navigating within the essay, hierarchy. Grendel Book! In my experience, this decision is science essay really one you should make in africans, conjunction with users. If you decide to heed this warning, select Quarter in essay, the Attributes pane and Mondays then, in the Properties window, change the AttributeHierarchyVisible value to science, False. Another warning appears on Close of Historical Theories: Challenging Narrative the hierarchy itself. In this case, the message cautions that a performance problem might arise because attribute relationships are missing between one or more levels of the hierarchy.

Attribute relationships are used by SSAS to optimize query performance and science aggregation design, to reduce the amount of grendel book storage required for a dimension, and to essay, speed up database processing time. Coca Vs Pepsi! Click the Attribute Relationships tab in science essay, the dimension designer. (Note that this tab is Readings of Conspiracy the Dominant available only essay if you're using Analysis Services 2008.) By default, all attributes relate directly to the key attribute, Date Key. Mondays Essay! To optimize the design by science essay, reassigning relationships, drag the Month object to the Quarter object, and then drag the Quarter object to the Year object. The relationship in grendel book, the designer now correctly represents the many-to-one relationship between each level from left to science essay, right, as shown in Figure 4 . Figure 4 Attribute Relationships. Next, add a hierarchy to the Product dimension called Categories that contains Category, Subcategory and Product from flies beast, top to bottom. Science Essay! After you create the hierarchy, you'll find you don't need to fix the attribute relationships between levels because the africans, foreign key relationships between the essay, tables are already defined in the DSV. Grendel Book! However, you can set the science, AttributeHierarchyVisible property value to False if you like. Cola Vs Pepsi! Now your project contains two dimensions that have hierarchies and attribute relationships correctly defined. Science! As you learn more about dimension design, you'll find there are several properties available for tuning performance and controlling specific behavior in the user interface. Also, you'll likely have several more dimensions in tu clothing online, more complex BI solutions. However, at this point, you have mastered the basics of science essay dimensions and have what you need to continue building a usable solution by pricing model, creating a cube.

Just as you use the Dimension Wizard to start the process of science essay defining your dimensions, you use the Cube Wizard to critical education, begin creating your cube. In Solution Explorer, right-click the Cubes folder, click New Cube and then Next on the Welcome page if applicable. On the Select Creation Method page, keep the default option. Essay! (You'd use the enslaved, other two options when you have a design you want to science, create manually or with a template, then have SSAS generate tables in lord, the data source based on that design, which you'd need to populate with data using Integration Services before you could browse your cube.) Click Next. On the Select Measure Group Tables page, choose Internet Sales and Reseller Sales and click Next. Science! (Measure group tables are synonymous with fact tables.) The wizard then displays all the thinking, numeric columns found in the selected measure group tables. In keeping with the simple approach you've taken thus far, clear all the measures by science, clearing the Measure check box at the top of the grendel book, page, and then select the following measures from science essay, each group (Internet Sales and beast Reseller Sales): Order Quantity, Total Product Cost and Sales Amount. You can also rename measures on this page. Just right-click on essay the measure name and beast type the new name, but make sure each measure name is unique. The measure name should be short, but not so short that it's cryptic.

Rename the measures in the Internet Sales group as follows: Internet Order Quantity, Internet Cost and Internet Sales. Likewise, rename the Reseller Sales measures as Reseller Order Quantity, Reseller Cost and Reseller Sales. Essay! Click Next. On the Select Existing Dimensions page, the wizard displays the dimensions you've already created. Click Next. Of The Flies Beast! If there are any tables in the DSV that are not yet referenced as a measure group table or by an existing dimension, the science, Select New Dimensions page will display to allow you to tu clothing, quickly add any other dimension you might need.

In this case, clear Internet Sales and science Reseller Sales as you don't need these tables created as dimensions. Flies Beast! (Although technically these are measure group tables, the science essay, presence of enslaved sales order numbers in the tables would allow you to create dimensions to support reporting or analysis of science sales by sales order number.) Click Next, rename the cube as Sales, and click Finish. Congratulations! You've successfully built a simple cube! The cube designer displays the capital asset pricing, measure groups and the dimensions added to the cube in essay, the panes on the left, and Readings and Fictional of Conspiracy the source DSV on the right, as shown in Figure 5 . Science Essay! Figure 5 Cube Designer Showing Measure Groups and Dimensions. Model! The first step to take after building a cube is essay setting the enslaved, FormatString property of science essay each measure to critical thinking, make it easier to view values in the cube browser. Science Essay! The quickest way to Close Readings of Historical Theories: Challenging, do this is to view the measures in essay, a grid. In the africans, toolbar of the cube designer, click the fifth button from the left to toggle from science essay, Tree View to Readings and Fictional of Conspiracy Challenging, Grid View. In Grid View, you can use the science essay, Ctrl key to pricing model, select several measures at once. First, select Internet Order Quantity and Reseller Order Quantity. In the Properties window, in essay, the FormatString dropdown list, select Standard.

Then select all remaining measures to set the FormatString property to africans, Currency. Essay! A powerful feature of SSAS is the Uptown in, ability to add calculations using the science, Multidimensional Expression language (MDX). If you can write formulas in Excel, you can create calculations in your cube using simple MDX expressions. For more complex expressions, you'll need to coca sales, spend some time learning about MDX through study and lots of practice. Remember that one of the science, design goals of this solution is to enslaved, measure profitability by sales channel. The cube now includes the measures you need to essay, calculate profitability: Internet Cost, Internet Sales, Reseller Cost and Reseller Sales. The difference between sales and cost is the gross margin, but that calculation provides absolute dollars, which isn't useful for enslaved, comparison between channels.

In addition to gross margin, you need to calculate the essay, gross margin percentage by education, dividing gross margin by essay, the sales amount. In the cube designer, click the Calculations tab, the third tab from the Kingston Jamaica, left. Science Essay! Then click the New Calculated Member button on asset pricing model the toolbar, the science essay, fifth button from the left. In the Name text box, type [Internet Gross Margin]. Kingston Jamaica Essay! The brackets are required when the name includes spaces.

In the essay, Expression text box, type [Internet Sales] - [Internet Cost], and then, in the Format String drop-down list, select Currency. Close Of Historical Narratives Theories: The Dominant Narrative! Now repeat these steps to add the calculations shown in Figure 6 . Figure 6 Adding Calculations to the Cube. Deploying an Analysis Services Database from BIDS. Essay! Thus far, although you have built the Readings Narratives Challenging the Dominant, objects necessary to create an SSAS database on the server, the database itself doesn't yet exist. Science! The design tasks you perform in BIDS produce XML files that must get deployed to the server. Once those files are deployed, you can process the database, which executes the online, commands necessary to build the storage structures defined by the XML files and to populate those structures by science essay, executing queries to grendel book, the specified data source. Science! Within BIDS, you start all of these activities by capital pricing model, right-clicking the project in Solution Explorer and then clicking Deploy.

The Deployment Progress window opens to show you each step it performs and whether that step succeeds or fails. After you have deployed a project, you can still make changes in the designers. Just deploy the project again as previously described to move your changes to science essay, the server and launch processing. If prompted that your database will be overwritten, click Yes if you're certain you're the only person making changes to the database. Sometimes the change you make won't trigger the online, command to process the database after deployment.

In this case, just right click the essay, dimension or cube you changed, then click Process, and in Kingston Jamaica Run. If you process a dimension using the Process Full option (which is required if you make a significant structural change to science, the dimension), you might also need to process the coca vs pepsi sales, cube. Science Essay! At each stage of development, as long as you have successfully deployed and Close and Fictional Narratives of Conspiracy Challenging the Dominant Narrative processed the solution, you can review your progress from the science, user's point of pricing model view. Science! In the cube designer, click the Browser tab. The pane on grendel book the left displays the essay, objects in the SSAS database in a metadata tree (shown in Figure 7 ). The top node of the tree is the cube. Enslaved! Expand the essay, Measures node and the folders it contains to view all of the measures available, and then expand the Order Date and Product nodes to africans, view the attributes in essay, these dimensions. Tu Clothing Online! Figure 7 Sales Cube Metadata Tree. You might be wondering why the cube contains Due Date, Order Date and Ship Date when you created only one Date dimension. These cube dimensions are called role-playing dimensions because they represent different logical versions of the same dimension. They appear automatically in the cube when you include the science, Date dimension because the Uptown in, fact table contains three different foreign key columns that all relate back to the single table on science essay which the coca vs pepsi, Date dimension is based to track the order, shipping and science due events separately. (If these role-playing dates aren't useful for analysis, you can delete any you don't need on the Cube Structure page of the designer.) To view cube data, you drag objects from the metadata tree to of the beast, the grid in the center of the science essay, designer.

Start by dragging Internet Sales to Readings and Fictional Narratives, the area labeled Drop Totals or Detail Fields Here. Then repeat this process to essay, add Internet Gross Margin Pct, Reseller Sales and Reseller Gross Margin Pct to the grid. With the online, results of this simple query now visible in the cube browser (as shown in science, Figure 8 ), you can see that Internet Sales are significantly more profitable than Reseller Sales. Figure 8 Query Results in grendel book, the Cube Browser. You can continue exploring these results by essay, dragging attributes to tu clothing online, the sections of the grid for rows, columns or filters, which collectively are called axes, or by dragging attributes and science essay measures off the grid. Of Historical Challenging Narrative! The process of adding objects to the axes to science, refine the coca, query is science essay called slice and dice by BI users and is a very fast way to tu clothing, query data without writing any code.

For example, to slice by science, year, drag Order Date.Year to the section labeled Drop Row Fields Here. In Kingston Jamaica Essay! Because Order Date.Year is a hierarchy (as indicated by the pyramid-shaped icon), you can drill down to slice by quarter by expanding one or more of the years. Similarly, you can dice by science, category by dragging Categories to coca cola vs pepsi, the columns axis above the science, measures. After you place objects on rows or columns, you can filter the grendel book, list of science essay items by clicking the Kingston, arrow in the caption. To remove Components from the science essay, grid, click the enslaved, arrow in the Category caption, clear the science, Components check box and click OK. Next, to coca cola, simplify the view, drag Internet Sales and Reseller Sales off the grid. You can now easily compare the essay, profitability of the Internet and Reseller channels by year and quarter and by enslaved, product category, as shown in Figure 9 . Essay! Figure 9 Profitability of Sales Channels by Year and Quarter and by Category.

By default, the in Jamaica, cube is accessible only by the server administrator. However, once your cube is science deployed to the Analysis Server, you can set permissions to grant users access to it, enabling them to use their favorite tool to browse the cube. Online! Microsoft Excel 2007 is a popular choice for science essay, interactive browsing, but you can also use Reporting Services to enslaved africans, distribute reports based on cube data, as I'll explain in science, a future article. Now that you've built a small database using the coca vs pepsi sales, sample AdventureWorks data, you should apply what you've learned to science essay, your own data by building a simple cube. If you have a simple design and coca sales a relatively small data set (containing less than a few million rows, for example), you can build a database without going to the effort of building and science essay maintaining a data mart. Just set up a data source view to asset pricing, query the source using named queries that structure the data as closely as possible to a star schema. Science Essay! When your data source has new data, you can simply do a full process of your SSAS database to tu clothing online, keep it current with your data source.

If you have more data to science, put into a cube, you should read more about SSAS in pricing, Books Online and essay other resources as this article explains only the very basic points you need to know to capital asset pricing, build a simple cube. Essay! For example, at grendel book the beginning of this article, I mentioned that one benefit of SSAS is aggregation management, but to explain more about that subject is beyond the scope of this article. You can learn more about aggregations and other advanced topics by science essay, reading Microsoft SQL Server 2008 Analysis Services Step by tu clothing online, Step (Microsoft Press, 2009), recently published by a good friend of mine, Scott Cameron, who started his business intelligence career with me many years ago. Now that you've built your first cube, keep enhancing your skills so that you can develop more advanced cubes and science take full advantage of the africans, Microsoft BI platform.

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Smalls-Mantey, Sample F31 Application and Summary Statement. We are truly indebted to the grantee who allowed us to post this outstanding F31 sample application to help the science essay next generation of investigators write applications. The F31 sample application below doesnt reflect the pricing complex layouts of the actual SF 424 PDF forms. Instead, weve provided a list of key fields from those forms and the applicants responses. Science? In addition, we redacted some applicant-specific information such as contact information, publications in press, and budget specifics. The text of this application is copyrighted. You may use it only for nonprofit educational purposes provided the document remains unchanged and the PI, the grantee organization, and NIAID are credited. Questions? Contact Critical? We also offer other outstanding Sample Applications. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms.

Here is the summary that eRA Commons generated at science, the start of the sample app, which reflects many key fields from that form.] Project/Performance Site Locations. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. Performance sites are in New York, Maryland, and the United Kingdom.] Research Related Other Project Information. [Our Web version of this sample application can't reproduce the pricing model layout of the actual PDF forms.

Here are the key fields from that form and the applicant's responses.] Understanding the basis of an science, immune response that controls infection or provides sterilizing immunity remains a major goal in the search for effective vaccines or immunotherapies for HIV. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in most recent vaccine trials. However, this approach has not been successful in tu clothing, containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an science, infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and use perforin to deliver granzymes which induce apoptosis in grendel book, the target.

We want to study ADCC in infected patients to understand the magnitude and characteristics of the best responses achieved by natural infection. Science Essay? First, we will compare ADCC mediated by the sera of a cohort of tu clothing patients using a granzyme B cytotoxicity assay developed in our lab. Based on these findings, we will select the sera of patients with the most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type. Science Essay? We will also evaluate whether ADCC is disparate from classical neutralization. Finally, we will use microscopy to examine the synapse between effectors, Abs, and targets. Asset Model? The outcome of this research will provide insight into science, the characteristics of Abs that mediate ADCC that are likely important goals in the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from tu clothing online lentiviral infection. We hypothesize that variations in ADCC activity of science essay sera are dictated by the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the potency of online sera of science HIV-infected individuals in ADCC. Aim 2 : Characterize the grendel book specificity and breadth of antibodies with ADCC activity.

Aim 3 : Characterize the structure and function of the target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography. Understanding the role of antibody-dependent cellular cytotoxicity (ADCC) in HIV could provide important insights for science essay, induction of this activity through vaccination. This project seeks to characterize the Abs that mediate ADCC and image the functional synapse formed by cellular components involved in ADCC with the goal of defining new goals for critical thinking education, the development of HIV vaccines and therapeutics. Gallo RC, Salahuddin SZ, Popovic M. Science? Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Lord Of The Flies? Science. 1984; 224: 500-3.

Barre-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, et al. Essay? Isolation of lord flies beast a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science. 1983; 220(4599): 868-71. Science? Organization WH.

Global summary of the AIDS epidemic, December 2007. Online? 2007 [cited 2009 September 18, 2009]; Hessell AJ, Hangartner L, Hunter M, Havenith CE, Beurskens FJ, Bakker JM, et al. Fc receptor but not complement binding is important in antibody protection against science essay, HIV. Nature. Enslaved? 2007; 449(7158): 101-4. RV144 Phase III HIV Vaccine Trial Press Release. 2009. Chung A, Rollman E, Johansson S, Kent SJ, Stratov I. The utility of science ADCC responses in HIV infection.

Current HIV research. Capital Asset Pricing? 2008; 6(6): 515-9. Kohl S, Starr SE, oleske JM, Shore SL, Ashman RB, Nahmias AJ. Human monocytemacrophage- mediated antibody-dependent cytotoxicity to herpes simplex virus-infected cells. Science Essay? J Immunol. 1977; 118(3): 729-35. Forthal DN, Landucci G. Cola Vs Pepsi Sales? In vitro reduction of virus infectivity by antibody-dependent cellmediated immunity. Journal of immunological methods. 1998; 220(1-2): 129-38. Stratov I, Chung A, Kent SJ. Robust NK cell-mediated human immunodeficiency virus (HIV)-specific antibody-dependent responses in HIV-infected subjects.

J Virol. 2008; 82(11): 5450-9. Gomez-Roman VR, Florese RH, Patterson LJ, Peng B, Venzon D, Aldrich K, et al. A simplified method for the rapid fluorometric assessment of antibody-dependent cell-mediated cytotoxicity. Journal of science essay immunological methods.

2006; 308(1-2): 53-67. Grendel Book? Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, et al. Lytic granule loading of CD8+ T cells is required for HIV-infected cell elimination associated with immune control. Immunity. 2008; 29(6): 1009-21. Vago L, Perna SK, Zanussi M, Mazzi B, Barlassina C, Stanghellini MT, et al. Loss of mismatched HLA in leukemia after stem-cell transplantation. N Engl J Med. Essay? 2009; 361(5): 478- 88. Ziegner U, Campbell D, Weinhold K, Frank I, Rutstein R, Starr SE. Deficient antibodydependent cellular cytotoxicity against sales, human immunodeficiency virus (HIV)-expressing target cells in perinatal HIV infection.

Clin Diagn Lab Immunol. 1999; 6(5): 718-24. Brunetta E, Fogli M, Varchetta S, Bozzo L, Hudspeth KL, Marcenaro E, et al. The decreased expression of Siglec-7 represents an early marker of essay dysfunctional natural killer cell subsets associated with high levels of HIV-1 viremia. Blood. 2009.

Ljunggren HG, Ohlen C, Hoglund P, Yamasaki T, Klein G, Karre K. Afferent and critical education, efferent cellular interactions in natural resistance directed against MHC class I deficient tumor grafts. J Immunol. 1988; 140(2): 671-8. De Maria A, Ferraris A, Guastella M, Pilia S, Cantoni C, Polero L, et al. Expression of HLA class I-specific inhibitory natural killer cell receptors in science, HIV-specific cytolytic T lymphocytes: impairment of specific cytolytic functions. Proceedings of the National Academy of asset model Sciences of the United States of America. 1997; 94(19): 10285-8. Ahmad R, Sindhu ST, Toma E, Morisset R, Vincelette J, Menezes J, et al. Evidence for a correlation between antibody-dependent cellular cytotoxicity-mediating anti-HIV-1 antibodies and prognostic predictors of HIV infection. J Clin Immunol. Science? 2001; 21(3): 227-33.

Lambotte O, Ferrari G, Moog C, Yates NL, Liao HX, Parks RJ, et al. Heterogeneous neutralizing antibody and grendel book, antibody-dependent cell cytotoxicity responses in HIV-1 elite controllers. Essay? AIDS. 2009; 23(8): 897-906. Dalgleish A, Sinclair A, Steel M, Beatson D, Ludlam C, Habeshaw J. Failure of online ADCC to predict HIV-associated disease progression or outcome in a haemophiliac cohort. Clin Exp Immunol. 1990; 81(1): 5-10.

Koup RA, Sullivan JL, Levine PH, Brewster F, Mahr A, Mazzara G, et al. Antigenic specificity of antibody-dependent cell-mediated cytotoxicity directed against science essay, human immunodeficiency virus in antibody-positive sera. J Virol. 1989; 63(2): 584-90. Doria-Rose NA, Connors M. Antibody-secreting B cells in HIV infection. Tu Clothing? Curr Opin HIV AIDS. 2009; 4(5): 426-30. Burke B, Barnett SW. Broadening our view of protective antibody responses against science, HIV.

Current HIV research. 2007; 5(6): 625-41. Sattentau Q. Avoiding the void: cell-to-cell spread of asset human viruses. Science Essay? Nat Rev Microbiol. 2008; 6(11): 815-26. Chamow SM, Zhang DZ, Tan XY, Mhatre SM, Marsters SA, Peers DH, et al. Asset Model? A humanized, bispecific immunoadhesin-antibody that retargets CD3+ effectors to essay kill HIV-1- infected cells. J Immunol. Capital Asset Model? 1994; 153(9): 4268-80.

Jolly C, Kashefi K, Hollinshead M, Sattentau QJ. HIV-1 cell to cell transfer across an Env-induced, actin-dependent synapse. J Exp Med. 2004; 199(2): 283-93. Groot F, Welsch S, Sattentau QJ. Efficient HIV-1 transmission from macrophages to T cells across transient virological synapses. Blood. 2008; 111(9): 4660-3. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D, Sattentau QJ. Virological synapsemediated spread of human immunodeficiency virus type 1 between T cells is sensitive to entry inhibition. J Virol.

2010; 84(7): 3516-27. Intellectual Support : My mentors Mark Connors and Quentin Sattentau are leading experts in their field and have mentored numerous people that have gone on to successful careers in science. This dual mentorship will teach me to aggressively approach science from two distinctive perspectives in my graduate career, something most PhD scientist dont experience until their first post-doctoral experience. While at NIH I will have received extensive support of science staff from Richard Siegel, Director of the NIH MD/PhD partnership program and Michael Lenardo, Director the NIH/Oxford/Cambridge Scholars Program. While at Oxford I will have the guidance of Lucinda Risius, Managing Director of the Graduate Pathology Program and I will also be assigned a departmental advisor, outside of my lab, with whom I can discuss my research Progress. Educational Resources : Through the FAES at NIH I have access to courses to improve lab specific skills and foundation knowledge.

I have already participated a Vaccine Development course. At Oxford I will take seminars at the Centre for Excellence in Teaching and Learning for training in teaching skills. I will also take courses pertinent to cola my research offered at science, the Dunn School of Pathology. Both the NIH and Oxford have libraries and IT staff that can aid me in scholarly research. Physical Resources : Both labs have sufficient funding to grendel book provided me any reagent/equipement need for the proposed research. Lab Space I am provided with a desk and science, personal desktop computer. Bench space is shared within the capital pricing lab. Safety In order to ensure our safety from hazardous blood samples we work under laminar flow hoods in a P2 facility with P3 practices. Samples We receive blood samples from science essay patients enrolled within our protocol. We also have access to the NIH blood blank for grendel book, normal human blood samples. As provided by Quentin Sattentau: Oxford University is held in high international regard for its strong teaching and research background and its unique and personal teaching style.

Within the University the Department of Pathology has an excellent reputation for graduate teaching, and has achieved 100% success in science essay, DPhil graduation within 4 years for the past 3 years. The Department has a strong graduate support structure, in which each student has a principal supervisor, the possibility of a cosupervisor, a departmental advisor, and a college advisor. The supervisor(s) will generally meet with the student multiple times a term, and each of the flies non-supervisory mentors will meet with the candidate approximately once a term and will file reports on the candidates progress. Science? The candidate will be expected to attend a series of high profile external seminars that take place once a week in term time, and a regular series of internal seminars from leading and more junior University and Departmental investigators and students. Adjoa will attend and participate in tu clothing, the weekly lab journal club, in which scientific papers are taken apart critically by members of the group, and a weekly laboratory meeting in which each lab member presents their ongoing research.

Adjoa will also be encouraged to science chat to other members of the department during tea lunch and coffee breaks, a longstanding Oxford institution! The Department has a fully equipped quiet library for literature research and other reading and lord flies beast, writing. Adjoa will have her own writing up space close to science her work bench, equipped with a portable computer and if required, an external screen. Adjoa will have full access to of the flies beast all core facilities, including the imaging suite with multiple light microscopes, a flow cytometry and sorting suite, an electron microscopy facility with full technical and scientific support. We also have strong connections with the biochemistry and science, chemistry departments and the Weatherall Institute of Molecular Medicine at the John Radcliffe Hospital for obtaining clinical samples and expertise. BD FACSAria IIu. BD Pathways 855. VICTOR Light luminescence plate reader. AutoMACS Pro Separator.

AutoMACS Classic Separator. Containment II/III Laboratory with 4 tissue culture hoods. Tissue culture. The laboratory has a new tissue culture facility comprised of 8 double hoods and associated incubators and ancillary equipment that is shared with three other virology groups. Virology. The department has a large, fully-equipped containment-III laboratory shared between two groups, with three tissue culture hoods and asset pricing model, associated incubators and ancillary equipment, in essay, which HIV-1 is grown and used to infect cells.

Flow cytometry. The department has a core flow cytometry facility with three BD analyzers (2 x FACScalibur 2 x LSR2) and two sorters. Molecular biology. Grendel Book? The Sattentau laboratory has all facilities for science, molecular biology carried out under containment levels I, -II and III. Light microscopy. The department has 5 confocal microscopes of varying sophistication and resolution, two of which are set up for capital asset model, live cell immunofluorescence work. The department has three recently acquired high-speed CCD cameras and associated microscopes for live cell immunofluorescence analysis. The Sattentau laboratory has a Zeiss Pascal confocal microscope and a new CCD camera set up with a Zeiss Axiovert 200 microscope for fixed and live cell work in science essay, the containment-III laboratory, allowing real time analysis of live, infected cells. Electron microscopy.

The department has two transmission electron microscopes and capital asset pricing, a scanning electron microscope, all of which are maintained and run as a central service facility by science, a skilled dedicated operator. Our collaborator in Oxford (Kay Grunewald) is located at the University Churchill campus, which is online 4 miles from the Pathology Department. He maintains and uses 3 microscopes set up for cryo-tomography as follows. 300keV FEG cryo-electron microscope FEI TF30 Helium Polara with GIF2002 imaging filter (2 x 2k CCD) 300keV FEG cryo-electron microscope FEI TF30 with 4 x 4kCCD 120keV cryo-electron microscope FEI T12 with 4 x 4k CCD. Research Related Senior/Key Person Profile. [Our Web version of this sample application can't reproduce the layout of the actual PDF forms. See below for the biographical sketch attachments.]

My interest in studying infectious diseases stems from the 2001 anthrax scare in the Washington D.C. area. I already had a strong interest in science but through my participation in the Gene Search Science Project at science, Catholic University the africans following summer I came to understand how medical research impacts treatment for essay, routine illnesses and also plays a role in national defense. Since middle school I have been preparing for a career in science. At Eleanor Roosevelt High School I was enrolled in the rigorous Science and Technology Program during which time in tu clothing, participated in the Gene Search Project and researched biochemical pathways that influence sickle cell anemia with Dr. Essay? William Winters at Howard University Hospital. This experience taught me how research and medicine are intertwined and inspired me to become a physicianscientist. At UMBC I was a member of the Meyerhoff Scholars Program and an HHMI Undergraduate Research Scholar where numerous lab experiences, seminars, rigorous classes, and multiple opportunities for scientific presentations prepared me for tu clothing online, a career as a scientist. My lab skills and understanding of a research career increased exponentially during my four years in Michael Summers lab. Not only did I learn about protein purification, gel electrophoresis, NMR, and ITC while studying MuLV RNA packaging, but more importantly of the time and discipline necessary for excellent research and the internal drive that keeps one focused. Working with Dr. Science? Nikola Pavletich I learned the value of immediately organizing my data so that others could analyze my results quickly.

In the tu clothing online lab of Tom Cech I had the responsibility of designing a new protocol to study telomerase assembly, a skill that has enabled me to science design a novel assay to investigate AIDS specific questions in critical thinking, my graduate studies. Essay? The medical education Ive received at Columbia has given me a unique perspective with which I approach scientific questions. Under the tutelage of Mark Connors and Quentin Sattentau, leaders in their fields on enslaved africans, nonprogressive HIV infection and viral transmission, my understanding of infectious disease and my capacity to pursue further research in these fields will be greatly enhanced. In addition, the courses, medical preceptorships, presentations, teaching, and career development activities I am undertaking in the forthcoming years make me an science, excellent P.I. for this grant. Academic and coca cola, Professional Honors. 2003 National Urban League Scholarship 2003 UMBC Meyerhoff Scholar 2003 UMBC Honors College 2003 HHMI Undergraduate Research Scholar 2004 NAD Distinguished Youth Award 2005 UMBC Minority Access to Research Careers Scholar 2005 Abby Rockefeller Mauze Award Charitable Trust Scholar for Outstanding Achievement 2006 Goldwater Scholar 2007 UMBC Top Senior Biochemistry Major 2007 Graduated Sum Cum Laude from UMBC 2007 NIH/Oxford/Cambridge Scholar 2010 International AIDS Vaccine Conference Scholarship 2011 Keystone Symposia Underrepresented Minority Scholarship. 2004 Omicron Delta Kappa Leadership Honor Society 2004 Golden Key International Honor Society, Sophomore/Alumni Director 2007 Phi Beta Kappa.

Dey A, York D, Smalls-Mantey A, Summers MF. Science? 2005. Composition and online, sequence-dependent binding of RNA to the nucleocapsid protein of Moloney murine leukemia virus. Biochemistry. 44(10), 3735-3744. Miyazaki Y, Irobalieva RN, Tolbert BS, Smalls-Mantey A, Iyalla K, Loeliger K, D'Souza V, Khant H, Schmid MF, Garcia EL, Telesnitsky A, Chiu W, Summers MF. 2010. Structure of a conserved retroviral RNA packaging element by NMR spectroscopy and cryo-electron tomography. J Mol Biol.

404(5), 751-772. Smalls-Mantey A, Winter W. 2003. Biosynthesis of science essay 2,3-diphosphoglycerate in Erythrocytes. Poster presentation at ERHS Science Fair and Prince Georges County Science Fair. March 2003.

Smalls-Mantey A, Dey, A, Phillip C, Summers MF. Capital Asset Pricing? 2004. Science? Characterization of tu clothing online High-Affinity Nucleocapsid Protein Binding Sites Within the science essay Moloney Murine Leukemia Virus RNA Packaging Signal. Poster presented at thinking, 2004 Annual Biomedical Research Conference for science essay, Minority Students, November 2004. Smalls-Mantey A, Min J, Pavletich N. 2005. Structural and Biochemical Studies of the Yeast Rad4-Rad23 Complex Bound to grendel book DNA.

Poster presented at Tri-Institutional Gateways to the Laboratory Program Symposium and 2005 Leadership Alliance. August 2005. Science Essay? Smalls-Mantey A, Miyazaki Y, Summers MF. 2007. Dimerization of Moloney Murine Leukemia Virus Nucleocapsid Protein Binding Sites Facilitates Binding.

Poster presented at 2007 UMBC Undergraduate Research and Creative Achievement Day. April 2007. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Keystone HIV Vaccines (X5) Meeting. March 2010. Coca Sales? Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu.

2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 Oxford Pathology Department Graduate Student Symposium. Science? June 2010. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. Thinking Education? 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells.

Poster presented at 2010 NIH/Oxford/Cambridge Scholars Research Colloquium. Smalls-Mantey A, Klein R, Doria-Rose N, Laub L, Rood J, Migueles S, Sattenatu. 2010. HIV-Specific Antibodies Mediate Rapid Antibody-Dependent Cellular Cytotoxicity Against Primary HIV-Infected CD4+ T Cells. Poster presented at 2010 International AIDS Vaccine Conference. September 2010. Smalls-Mantey A. 2010.

Antibody-Dependent Cellular Cytotoxicity and the Control of HIV. Talk given at Frontiers in science, Biomedicine NIH/Oxford/Cambridge Scholars Program student presentation series. Of The Flies? November 2010. [Outside research support redacted from sample.] A. Personal Statement Regarding Project.

Currently HIV-specific ADCC is considered one of the science essay very top priorities in the field of tu clothing online HIV-specific immunity. Our laboratory has assembled the best techniques and science, reagents in the field to tu clothing online carry out this project. We have extensive experience in HIV-specific antibodies and science, cytotoxicity. In addition we have a long track record of training students to prepare them for a carreer in biomedical research. 1985 - 1986 Intern in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1986 - 1988 Resident in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts 1988 1989 Chief Resident in Pediatrics, The Boston Floating Hospital, New England Medical Center, Boston, Massachusetts July 1989 - October 1989 Medical Staff Fellow, Respiratory Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD October 1989 - June 1993 U.S. Public Health Service, Research Associate (C.O.), Respiratory Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and lord, Infectious Diseases, National Institutes of Health, Bethesda, MD July 1993 - June 1994 U.S. Public Health Service, Clinical Associate, Clinical Center, National Institute of science Allergy and Infectious Diseases, National Institutes of lord of the Health, Bethesda, MD January 1994 - June 1994 Clinical Fellow in Infectious Diseases, The Childrens Hospital of Philadelphia, Philadelphia, PA. June 1994 - December 2004 U.S. Public Health Service, Medical Officer, National Institute of Allergy and Infectious Diseases, Clinical and essay, Molecular Retrovirology Section, Laboratory of africans Immunoregulation, National Institutes of essay Health, Bethesda, MD December 2007- Present Chief, HIV-Specific Immunity Section, Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD. Recent Honors and of the flies beast, Editorial and Review Board Activities. 2009 - U.S.

Public Health Service Outstanding Service Medal 2009 - present Vaccine Production Development Team Scientific Advisory Board, University of Pennsylvania 2009 - present HIV Enterprise Host Genetics and HIV Diversity Working Group 2010 - present Editorial Board, Journal of essay Virology. C. Peer-Reviewed Publications. Migueles SA, Sabbaghian MS, Shupert WL, Bettinotti MP, Marincola FM, Schwartz D, Sullivan J, and Connors M. Grendel Book? HLA B*5701 is highly associated with restriction of science virus replication in a subgroup of HIV infected long term nonprogressors. PNAS 2000; 97(6): 2709-2714. Migueles SA, Laborico AC, Shupert WL, Sabbaghian MS, Rabin R, Hallahan C, Van Baarle D, Kostense S, Miedema F, McLaughlin M, Ehler L, Metcalf J, Liu S, and of the flies beast, Connors M. HIV-specific CD8+ T cell proliferation is coupled to perforin expression and science essay, is maintained in grendel book, nonprogressors. Nature Immunol 2002; 3(11); 1061-1068.

Migueles SA, Laborico AC, Imamichi H, Shupert WL, Royce C, McLaughlin M, Ehler L, Metcalf J, Liu S, Hallahan CW, and Connors M. The differential ability of HLA B*5701+ long-term nonprogressors and progressors to restrict HIV replication is not caused by loss of recognition of autologous viral gag sequences. Essay? J Virol 2003; 77(12); 6889-6898. Iyasere C, Tilton JC, Johnson AJ, Younes S, Yassine-Diab B, Sekaly RP, Kwok WW, Migueles SA, Laborico AC, Shupert WL, Hallahan CW, Davey RT Jr., Dybul M, Vogel S, Metcalf J, and Connors M. Diminished proliferation of HIV-specific CD4+ T Cells is associated with diminished IL-2 production and recovered by exogenous IL-2. Online? J Virol 2003; 77(20); 10900-10909. Tilton JC, Luskin MR, Johnson AJ, Manion M, Hallahan CW, Metcalf JA, McLaughlin M, Davey RT Jr., and Connors M. Changes in science, paracrine IL-2 requirement, CCR7 expression, frequency, and cytokine secretion, of HIV-specific CD4+ T cells are a consequence of antigen load. Online? J Virol 2007; 81;2713-2725. Migueles SA, Osborne CM, Royce C, Compton AA, Joshi RP, Weeks KA, Rood JE, Berkley AM, Sacha JB, Cogliano-Shutta NA, Lloyd M, Roby G, Kwan R, McLaughlin M, Stallings S, Rehm C, OShea MA, Mican J, Packard B, Komoriya A, Palmer S, Wiegand AP, Maldarelli F, Coffin JM, Mellors JW, Hallahan CW, Follman DA and Connors M. Lytic Granule Loading of CD8+ T-Cells is Required for HIV-Infected Cell Elimination Associated with Immune Control. Immunity 2008; 29 (6): 1009-1021.

Migueles SA, Weeks KA, Nou E, Berkley AM, Rood JE, Osborne CM, Hallahan CW, Cogliano-Shutta NA, Metcalf JA, McLaughlin M, Kwan R, Mican JM, Davey RT Jr, Connors M. Defective human immunodeficiency virus-specific CD8+ T-cell polyfunctionality, proliferation, and science essay, cytotoxicity are not restored by antiretroviral therapy. J Virol 2009 83(22): 11876-11889. Doria-Rose NA, Klein R, Daniels M, O'Dell S, Nason M, Lapedes A, Bhattacharya T, Migueles S, Wyatt RT, Korber BT, Mascola JR, Connors M. Breadth of HIV-Specific neutralizing activity in sera: clustering analysis and association with clinical variables. J Virol 2009 84(3): 1631-1636. Doria-Rose NA and Connors M. Antibody-secreting B cells in HIV infection. Curr Opin HIV AIDS 2009 4:426-430 Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou T, Schmidt SD, Wu L, Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, Nason M, Doria-Rose N, Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ and Mascola JR. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1.

Science 2010 329:856-861. Africans? Migueles SA and Connors M. Long-term nonprogressive disease among untreated HIV-infected individuals: clinical implications of understanding immune control of science essay HIV. JAMA 2010 304:194-201. [Redacted publication in press] The research direction of the HIV-Specific Immunity Section of the Laboratory of Immunoregulation is focused upon mechanisms of immunity to pricing model HIV. The laboratory is staffed by PhD and MD scientists with extensive experience in the humoral or cellular immune response to essay HIV. Tu Clothing? In addition we have close collaborations with investigators at the NIAID Vaccine Research Center in the area of structure-function relationships of science essay HIV-specific antibodies. Our laboratory is well equipped with multi-laser flow cytometers, automated magnetic sorters, and imaging equipment to carry out this project.

Our laboratory is supported by the research budget of the grendel book Division of Intramural Research. Over the science past 4 years we have received supplemental funding from the Intramural AIDS Targeted Antiretroviral Program (IATAP). Flies? This will continue for science, the next 2 years. We have also received funding from the Office of AIDS Research for work on the humoral response to HIV. Positions and Employment.

1985 - 1992 Lecturer, Department of Genito-Urinary Medicine, University College London, UK 1989 - 1991 Visiting scientist, Howard Hughes Medical Institute, Columbia University, New York, NY 1992 - 1998 Staff scientist (Director of Research), Centre dImmunologie, Marseille, France 1999 - 2003 Reader, Section of Infectious Disease, Imperial College London, London UK 2003 - Professor of immunology, Department of Pathology, University of Oxford, Oxford UK. Other Experience and Professional Memberships. 1995 - Member, American Association for the Advancement of Science 1998 - Member, American Society for coca cola vs pepsi sales, Microbiology 1998 - Member, British Society for Immunology, UK 1998 - Member, Society for General Microbiology, UK 2005 - Member of the Pasteur Institute Paris France Scientific Evaluation Committee. 2005 - Member of the Vaccine Research Center (VRC) NIH Board of Scientific Councilors 2007 - Member of BMGF Vaccine Monitoring Centre SAB 2008 - Member of NIH HIVRAD/Novartis Neutralizing Antibody Vaccine Consortium SAB 2009 - Member of essay study section for Swedish Program for International Development 2010 - Member of NIH study section for the B cell Immunology Partnership Program for HIV-1 Vaccine Discovery (U19) C. Grendel Book? Selected Peer-Reviewed Publications. Most relevant to the current application. Martin N, Welsch S, Jolly C, Briggs JA, Vaux D and Sattentau QJ. (2010) Virological synapse-mediated spread of human immunodeficiency virus type-1 between T cells is sensitive to entry inhibition. J. Virol. 84: 3516-3527. Gonzalez N, Bermejo M, Calonge E, Jolly C, Arenzana-Seisdedos F, Pablos JL, Sattentau QJ and essay, Alcami J. Grendel Book? (2010) SDF-1/CXCL12 production by mature dendritic cells inhibits the essay propagation of X4-tropic HIV-1 isolates at tu clothing, the dendritic cell-T-cell infectious synapse.

J Virol. 84:4341-51 Groot F, Welsch S and Sattentau QJ. (2008). Efficient HIV-1 transmission from macrophages to science T cells across transient virological synapses. Blood 111: 4660-4663. Sowinski S, Jolly CJ, Berninghausen O, Purbhoo, MA, Chauveau A, Kohler K, Oddos S, Eissmann P, Brodsky FM, Hopkins C, Onfelt B, Sattentau QJ and Davis DM (2008).

Membrane nanotubes physically connect T cells over long distances presenting a novel route for thinking education, HIV-1 transmission. Nature Cell. Biol. 10: 211-219. Jolly C and Sattentau QJ (2007).

HIV-1 assembly, budding and cell-cell spread in science essay, T cells takes place in tetraspanin-enriched plasma membrane. J. Virol. 81:7873-7884 Jolly C and Sattentau QJ (2007). Adhesion molecule interactions facilitate human immunodeficiency virus type-1-induced virological synapse formation between T cells. J. Virol. 81: 13916-13921. Jolly C, Mitar I and Sattentau QJ (2007). Requirement for an intact actin and tubulin cytoskeleton for efficient HIV-1 assembly and spread.

J. Virol. 81: 5547-6 Jolly C and Sattentau QJ. Human Immunodeficiency virus type-1 virological synapse formation in T cells requires lipid raft integrity. J. Virol. 2005 79: 12088-12094.

Jolly C, Kashefi K, Hollinshead M and education, Sattentau QJ (2004) HIV-1 cell-to-cell transfer across an Env-induced, actin-dependent synapse. J. Exp Med. 199: 283-193. Additional recent publications of important to the Field (in chronological order) Wegmann F, Krashias G, Luhn K, Laamanen K, Vieira S, Jeffs SA, Shattock RJ and science essay, Sattentau QJ (2011) A novel strategy for inducing enhanced mucosal HIV-1 antibody responses in an anti-inflammatory environment. PlosOne 6: e15861. Kong L, Sheppard N, Stewart-Jones G, Robson CL, Chen H, Xu X, Krashias G, Bonomelli C, Scanlan CN, Kwong PD, Jeffs SA, Jones IM, Sattentau QJ (2010) Expression system-dependent modulation of HIV-1 envelope glycoprotein antigenicity and immunogenicity. J. Mol. Biol. Aug 25th epub. Krashias G, Simon K, Wegmann F, Kok WL, Ho LP, Stevens D, Skehel J, Heeney JL, Moghaddam AE and Sattentau QJ. (2010).

Potent adaptive immune responses induced against HIV-1 gp140 and influenza virus HA by a polyanionic carbomer. Vaccine 28: 2482-2489. Zanetti G, Briggs JAG, Grunewald K, Sattentau QJ and Fuller SD. SIV spike glycoprotein structure in situ determined by cryo-electron tomography. Grendel Book? Plos Pathogens August 2 (8): e83 Sheppard NC, Bates AC and Sattentau QJ (2007). A functional human IgM response to HIV-1 Env after immunization with NYVAC-HIV C. Science? AIDS 19: 524-527. [Outside research support redacted from sample.] PHS Fellowship Supplemental Form, Research Plan. [Our Web version of coca this sample application can't reproduce the layout of the actual PDF forms. Science Essay? The attachments are included below. In the original version, these attachments met the lord page limits.]

Understanding the basis of an science, immune response that controls infection or provides sterilizing immunity remains a major goal in the search for grendel book, effective vaccines or immunotherapies for HIV. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the primary goal in most recent vaccine trials. However, this approach has not been successful in containing viral replication in essay, vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. Enslaved? In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and science essay, an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the capital asset Ab and use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in essay, infected patients to understand the magnitude and characteristics of the best responses achieved by natural infection. First, we will compare ADCC mediated by the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on these findings, we will select the sera of patients with the enslaved most ADCC, generate monoclonal Abs (mAbs), and characterize the mAbs based on epitope specificity, affinity, potency, breadth, IgG isotype, and Fc type.

We will also evaluate whether ADCC is disparate from science essay classical neutralization. Finally we will use microscopy to examine the synapse between effectors, Abs, and targets. The outcome of this research will provide insight into the characteristics of Abs that mediate ADCC that are likely important goals in tu clothing, the design of HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to essay mediate protection from grendel book lentiviral infection. Science? We hypothesize that variations in ADCC activity of sera are dictated by critical, the amount, specificity, and subclass of HIV-specific antibodies. Aim 1 : Characterize the science essay potency of critical sera of HIV-infected individuals in ADCC. In ADCC, Abs bind viral epitopes that are presented by infected CD4+ T cells. Essay? NKs expressing an Fc receptor bind the Fc domain of the Ab and use perforin to deliver granzymes to the HIV-infected cell.

Subsequently, granzymes induce apoptosis within the cell. Grendel Book? Our lab has developed a flow cytometric assay that measures granzyme B delivered to science essay an HIV-infected CD4+ target T cell. We will classify ADCC by the percent of target cells receiving granzyme and the elimination of targets as defined by residual percent of targets expressing p24, HIV capsid. Compare the serum of HIV+ individuals with various rates of coca sales progression and viral loads to science determine which contain Abs capable of mediating the highest levels of ADCC. Compare the of the flies ADCC and neutralizing activity of science patient sera. Aim 2 : Characterize the specificity and breadth of antibodies with ADCC activity. Our laboratory has panels of critical thinking NAbs derived from patients with known serum neutralizing or ADCC-mediating activity. Determine whether recognition of specific epitopes is required for science, ADCC. Define the breadth of the polyclonal sera by its ability to mediate ADCC in CD4+ T cells infected by different clades of HIV.

Titer serum total IgG, IgG1, and IgG3 binding infected CD4+ T cells. Aim 3 : Characterize the grendel book structure and function of the target-effector synapse. Using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography, we will examine the synapse formed between NK and other cells with potential ADCC activity (macrophages and neutrophils) and infected target cells. We will specifically investigate: The structure of a functional ADCC synapse. The kinetics of ADCC function in real time and science, its relation to africans antibody type and specificity. A role for antibody-dependent cell-mediated phagocytosis (ADCP) in elimination of HIV-infected cells. d. Receptors and science essay, effector molecules central to ADCC activity against HIV infected cells. Since being identified as the causative agent of AIDS in 1983, HIV infection has reached epidemic proportions with an estimated 2 million people worldwide dying from AIDS each year 1- 3. Pricing? There is more public awareness about how to prevent HIV infection but with the growing incidence of HIV infection and the limitations of science current antiretroviral therapy, there is an imperative need for asset pricing model, a vaccine.

One of the most critical areas of research on vaccines for HIV is the search for correlates of immunity. At present the only method for determining the potential benefits of a vaccine with regard to protection from infection or lowering of science essay viral load upon infection remains very large clinical efficacy trials. Determining correlates of immunity would tremendously accelerate the development process by permitting some estimation of grendel book clinical efficacy prior to a Phase III trial. Science? Reliable correlates would permit screening of vaccine candidates at a much earlier point in coca cola vs pepsi sales, development with smaller numbers of participants. Over the past 20 years, research into science essay, correlates of tu clothing online immunity to science essay HIV have largely focused on grendel book, CD8+ cytotoxic T lymphocytes (CTLs) or neutralizing antibodies (NAbs). In addition to neutralization, antibodies (Abs) may fight viral infections by antibody-dependent cell-mediated phagocytosis (ADCP) or by antibody-dependent cellular cytotoxicity (ADCC). In ADCC, Abs bind viral epitopes presented on infected target cells and Fc? receptor-bearing effector cells such as natural killer cells (NKs) recognize and bind the Fc? region of essay these Abs. Upon cross-linking of the Fc? receptor, NKs release perforin that punctures the targets cell membrane and deliver proteases such as granzyme B (GrB) which induce apoptosis. ADCC has become a major focus within the field following two important studies highlighting its potential importance in providing immunity to HIV.

A recent study by Burton and colleagues demonstrated that neutralizing Abs with mutated Fc? receptor binding regions were less effective in protecting macaques from HIV whereas Abs with impaired complement binding conferred protection to tu clothing online macaques, suggesting that ADCC may be more important in preventing infection than neutralization or complement activation alone 4. Prior to this study, it was widely accepted that infection of a single cell would lead to integration and chronic infection. However, because ADCC mediated effects are on virus infected cells and essay, not free virions, this study provided indirect evidence and proof of the concept that ADCC can eliminate lentivirus-infected cells in enslaved africans, vivo. In addition, some recent data in human efficacy trials has also greatly increased interest in ADCC as an important component of vaccine induced immunity to HIV. In September 2009 data from the phase III RV144 (Thai) vaccine trial were released and showed that the prime-boost combination of ALVACR HIV and AIDSVAXR B/E reduced HIV infection. This vaccine regimen was known to not elicit a strong cellular or NAb response. Science Essay? It does, however, elicit binding antibodies that have suggested to the field that the protective efficacy is likely mediated through ADCC 5. Our laboratory is part of critical a large international effort to characterize ADCC antibodies from this trial in order to determine whether there were higher ADCC-mediating antibodies in uninfected vaccinees compared to those that were infected. Although HIV-specific ADCC is now considered an essay, extremely important part of vaccineinduced immunity several basic questions regarding the nature of an effective ADCC response have not been addressed. For example, the number, affinity, and specificities of grendel book antibodies necessary to science mediate ADCC remains poorly understood. Our laboratory has now developed assays that can accurately measure ADCC against vs pepsi sales, HIV-infected cells. We also have assembled panels of essay monoclonal antibodies from individual patients that permit a characterization of individual antibodies that mediate HIV-specific ADCC.

One of the of the flies beast initial goals of this project was to essay adapt our current killing assay to provide a more robust assay for HIV-specific ADCC. Past studies of ADCC have been hampered by the availability of assays that are only partially quantitative. Coca Vs Pepsi? There are four primary methods that people use to science essay measure ADCC 6. The most prevalent is the chromium release assays (CRA), which is highly variable, does not specifically account for the mechanism of killing, and is insufficiently quantitative 7. Enslaved Africans? The ADCVI assay incubates targets, serum, and effectors for science, 24 hours and the supernatant is applied to coca sales the B95-8 cell line for 5-7 days. The endpoint is reduction of viral infection of the B95-8 cells 8. This method measures inhibition of viral production and subsequent infection but its disadvantages lie in the length of the assay and failure to measure killing of infected cells. Another assay is the intracellular cytokine staining ADCC (ICS-ADCC) assay measures IFN-?, the primary cytokine responsible for the antiviral activity of many cells including NKs, CTLs, Th1, and dendritic cells 9. The advantages of this assay are that multiple effector functions can be measured and all cells and Abs are from the science same patient, but it is still a proxy that does not measure HIV-infected cell killing.

The rapid and tu clothing online, fluorometric antibody-dependent cellular cytotoxicity (RFADCC) assay directly measures cell killing but requires whole protein 10. While use of essay protein pulsed targets is a sensitive method, it likely places artificially high amounts of protein on the surface of infected cells, potentially overestimating the efficacy of ADCC. Recently our lab has developed a highly quantitative flow cytometric assay to measure GrB cytotoxicity using primary HIV-infected CD4+ T cells as targets and grendel book, autologous NKs as effectors. This is an adaptation of an assay for measuring the cytotoxic activity of CTLs developed in our lab11. This assay measures the essay cleavage of a GrB substrate, and eliminates background cell death using a Live/Dead stain to produce accurate quantitative and highly reproducible results. In brief, CD4+ T cells are stimulated for 3 days and HIV infected. Autologous, negatively selected NKs are used as effectors. Effectors and targets are labeled with surface dyes so that they can be discriminated. NKs, targets and serum are incubated in the presence of a granzyme cleavable fluorescent substrate.

Granzyme delivery to the target cell and grendel book, elimination of science essay HIV-infected targets can then be measured by flow cytometry (Figure 1). Cellular GrB content. GrB cleaves GranToxiLux substrate (FITC), indicating the amount of GrB present within a target cell (Pacific Blue). Elimination of HIVSF162-infected cells. Live targets are stained for intracellular HIV capsid, p24 (PE). Grendel Book? Some infected cells show downregulation of essay CD4 (PE-Cy7). Critical Thinking Education? A reduction in the number of p24 stained cells is represented as Infected Cell Elimination (ICE) and is an indication of essay cell death. In the tu clothing example above, ICEInfected+Effectors+Serum = [(58.2-25.8)/58.2] x 100 = 54.9%.

There are several advantages to this assay over older assays that use mismatched cell lines. Because NKs lyse target cells in essay, the context of MHC mismatch and cause nonspecific killing, autologous cells are used in our assay to enslaved reduce background. 12. In addition, because NK function is diminished in HIV-infected patients, we use the NK cells and CD4+ T cell targets from uninfected patients in our assay as a readout for the effectiveness of essay a given serum sample in mediating ADCC 13, 14. We observed variations in NKs from grendel book normal individuals in their ability to mediate ADCC and choose donors that provide optimal signal to science essay noise ratio that we will use to compare the serum of HIV+ individuals (Figure 2). The targets used in this assay are likely closer to tu clothing online the in vivo situation given autologous HIV-infected targets are used in science, the place of antigen pulsed cell lines.

Other advantages include our assays ability to measure the mechanism of online ADCC (granzyme delivery) and science essay, the outcome (death of infected cells) (Figure 2). MHC-1 is down-regulated in virally infected cells, and the loss of MHC-mediated killer inhibitory signals permit NKs to kill a cell 15, 16. We control for this by measuring the granzyme content of grendel book targets that are incubated only with effectors and essay, sera from lord beast HIV-uninfected volunteers. For a positive control we are using a novel anti-CD3 Ab that will mediate ADCC to all CD3-bearing targets (Figure 3). With this Ab we have already demonstrated (1) the sensitivity and science essay, dynamic range with which we are able to measure ADCC, (2) Abs mediate ADCC in a dose dependent manner, and (3) GrB delivery is proportional to cell death. Aim 1: Characterize the magnitude of ADCC in HIV infected individuals . Rational : There is considerable controversy regarding the magnitude of ADCC activity in sera from HIV+ individuals and how it correlates with disease progression.

It has been suggested that sera from elite controllers exhibit greater ADCC activity which correlates with CD4+ T cell count 17, 18 while others have sighted no correlation between ADCC and disease progression 19, 20.We will compare the of the beast serum of HIV+ individuals with various rates of progression and essay, viral loads to determine which contain Abs capable of mediating the critical education highest levels of ADCC. We will also compare the ADCC and neutralizing activity of patient serum to evaluate whether or not ADCC and neutralization always occur together in polyclonal sera which is yet to be determined 21, 22. Methods : Over the past several years, in collaboration with the Vaccine Research Center (VRC) our laboratory has focused upon recruiting HIV-infected patients with broadly crossneutralizing antibodies. We have extensive clinical data and neutralization data for all of the patients. With the data we generate from our ADCC assay we are able to compare multiple indicators of immunity with ADCC. Expected Outcomes/Interpretation : Based on essay, in vitro and asset, passive transfer macaque studies, we expect LTNPs to exhibit greater ADCC than progressors. Our current knowledge attributes LTNP delayed progression to superior CTL cytotoxicity but our data may show ADCC also to be a significant factor 11. Potential Complications : We do not foresee any difficulties in accomplishing this objective. Aim 2: Characterize MAb from serum identified as having potent ADCC activity with regard to essay specificity. Rational/Methods : HIV-infected cells express Envelope (Env) protein on their surface which is recognized by enslaved, NAbs and presumably by science, ADCC-mediating Abs.

We will use standard Nabs (including b12, 2F5, 2G12, 447-52D, 17B, and coca sales, VRC01) and patient derived MAbs to science essay probe specificities required for grendel book, ADCC. An ideal HIV vaccine would be able to protect against diverse virus strains so it is important to science determine whether Abs raised against tu clothing online, one clade of virus would be able to essay mediate ADCC in lord of the, CD4+ T cells infected by science essay, different clades. We will use multiple strains of virus from clade A/E, B, and C to enslaved africans infect CD+ T cells and compare killing of target cells in the presence of essay serum pooled from clade B HIV+ patients. IgG1 and IgG3 are the predominant IgG isotypes with roles in ADCC. It is possible that variations in ADCC activity of sera are mediated by variations in africans, IgG subclass. Science? We will use infected CD4+ T cells, apply sera with various levels ADCC at the same dilutions used in our killing assay, and measure total bound IgG, IgG1, and online, IgG3 by flow cytometry. We believe flow cytometry is science essay a better way of measuring titers of binding Abs as compared to ELISA because Abs will recognize all possible epitopes on vs pepsi, the surface of an infected cell in their natural conformation. Expected Outcomes/Interpretation : Determining the association between ADCC activity and IgG subclass may provide insights into the necessity of eliciting a specific class of IgG via particular antigens and adjuvants. Potential Complications : We now have robust assays for subclass binding and do not expect complications with this portion of the project.

Aim 3: Evaluation of the target-effector synapse. Rational : Using microscopy to study ADCC is science essay important because it will help us define structural and lord of the beast, kinetic parameters and will relate structure to function. Multiple mechanisms for cell-to-cell spread of HIV have been described but few have described inhibition of viral transmission or targeted cell death 23. To date there has been no study using microscopy to essay investigate ADCC in HIV using NKs or macrophages as effectors and CD4+ T cells as targets. In 1994 one group designed a bispecific antibody with anti-CD4 and anti CD3 heavy chains that would redirect CD8+ T cells to grendel book kill HIV infected targets 24.

With fluorescent microscopy they observed after 1 hour of incubation that 24% targets had formed conjugates with CD8+ T cells although many more cells had lysed. Science? At 2 hours 50 - 80% of targets were conjugated and undergoing lysis and by 24 hours all targets had been lysed. However, the bispecific antibody may not be representative of ADCC with naturally occurring HIV-specific antibodies. In addition, the structure of the NK to HIV-infected target synapse was not examined. Methods : The lab of Quentin Sattentau has focused on examining the synapse of cells involved in coca cola vs pepsi sales, HIV infection. They were the first to demonstrate HIV-1 spread between T cells via virological synapses and nanotube structures for viral transmission between CD4+ T cells 25. They have recently demonstrated that macrophages can infect CD4+ T cells every 6 hours and that cell-tocell transfer of virus is science 10-fold more efficient than free virus spread 26.

To study the kinetics of pricing model ADCC in real time we will use time-lapse laser scanning confocal microscopy (time-lapse LSCM). Essay? Cell populations will be negatively selected from PBMCs using magnetic cell sorting. HIV-infected CD4+ T cells will be mixed with patient serum or Abs of multiple specificities (see Aim 2) and NKs to flies evaluate ADCC; or neutrophils/monocytes to science evaluate antibody-dependent cell-mediated phagocytosis (ADCP) and applied to poly-L-lysine coated cover slips. Cover slips will be fixed in paraformaldehyde, quenched with NH4Cl, permeabilized with Triton-X, and stained for granzyme B, IgG, CD4/CD16/CD14 with fluorescent probes. After mounting, images will be acquired every minute for 6 hours using a Zeiss Pascal Axivert 200M scope.

We will use transmission electron microscopy and lord flies, tomography (TEM) to science generate higher resolution static images of the ADCC functional synapse. For TEM targets, serum, and effectors will be incubated for approximately 1 hour, fixed in paraformaldehyde and glutaraldehyde, and coca, stained with osmium tetroxide. Samples will be embedded in epoxy resin, cut, and imaged using a 300kV FEI Technai TF30 27. If necessary cryo-EM will also be used for structural studies with the science essay aid of lord Oxford collaborator Kay Grunewald. These experiments will be completed using the pathology departments core bioimaging facility and I will take departments Advanced Light Microscopy and Electron Microscopy course in science, preparation for this research. Expected Outcomes : We expect to elucidate the kinetic and functional relationship between ADCC components and answer questions such as what density of Env epitopes are needed to recruit effectors, how many effectors are recruited per critical, target. Science Essay? We do not foresee any difficulties in accomplishing this objective. The results of these studies will include novel MAb with possible therapeutic value; a new tool for the study of cola ADCC with potential utility for HIV vaccine trials; and increased understanding of basic HIV immunology. In the wake of the RV144 trial, ADCC has become a major focus in the field of HIV-specific immunity. We are very well positioned to carry out these important studies because of essay our large collection of cells, sera, and detailed clinical data from patients that are part of the NIAID HIV clinic, and both labs extensive experience in this area.

Samples are collected under the NIAID IRB-approved protocol 02-I-0086. Protections for human subjects are in accordance with NIAID Division of Intramural Research policies. Phlebotomy may cause some discomfort, and occasionally some bleeding or bruising at the site. On rare occasions, people faint while having blood drawn. Blood drawing will not exceed 150 milliliters (30 teaspoonfuls) on africans, any day or visit. In the course of science any time frame during this study, the total amount of blood will not exceed 10.5mL/kg, or 550 milliliters over any 8-week period, which is within the safety guidelines for blood drawing practiced at the Clinical Center. Like standard phlebotomy, leukapheresis is occasionally associated with local bleeding and tu clothing, bruising at the site of needle insertion.

Lightheadedness can also occur, and people faint on rare occasions. Occasional tingling in the arms and legs can occur with the procedure and science, can usually be relieved by capital asset, chewing a calcium-rich tablet that the staff will provide. In addition, on very rare occasions (less than 1 in science, every 1,000 times), malfunction of the leukapheresis machine can result in blood loss of up to 450 milliliters. Trained medical personnel who are equipped to deal with any of these potential problems will always be on hand during the procedure. Inclusion of Women and asset, Minorities. Gender, Ethnicity and Race Consideration. This study involves techniques and procedures that are currently approved for various indications. Male and science, female adult patients (18 years of age or older) will be enrolled without preference to gender, race or ethnicity from the pool of of the flies patients that receive care in Clinics 8 at the NIH Clinical Center. Patients will be recruited from outside the NIH community by advertisements posted in the Clinical Center, the NIAID web site, and advertised in other media. We will attempt to facilitate minority recruitment using existing NIAID community outreach efforts involving local clinics in the Washington, DC metropolitan area. In adult HIV-infected patients planning to interrupt HAART, patients will be fully counseled prior to entry into essay, the study as to the potential risks of discontinuing HAART therapy and the potential benefits of continuing HAART therapy, in thinking, the setting of a documented virologic and/or immunologic response.

Patients who do not fully comprehend these potential risks of HAART cessation, in the opinion of the study team, will not be offered participation in the study. Patients stopping HAART therapy will be monitored closely for virologic and immunologic relapse and will be offered therapy, based upon science the algorithm for safety outlined above. Target enrollment for pricing, this study is 300 individuals and is currently ongoing. Inclusion of Children (and Pregnant Women) Inclusion of science Children and Pregnant Women. This study will be confined to tu clothing HIV-infected adults greater than or equal to 18 years of science essay age.

As an observational study there is no therapeutic benefit or clinical intervention resulting from lord flies beast participation in science, this trial, thereby children are not being denied the potential benefits of a research intervention. Grendel Book? We also do not have the historical research database and in vitro findings on HLA linkages with LTNP in the pediatric population to make valid comparisons within that cohort. The potential risks of withdrawing therapy in essay, minors have not been studied and therefore may present unknown additional risks to that study population. Comparable units specializing in pediatric HIV are available at grendel book, the NIH to address this topic, should preliminary results from this study warrant further evaluation in essay, the pediatric population. The safety of grendel book discontinuing antiretroviral therapy has not been established during pregnancy and nursing, and current guidelines continue to advise HIV-positive women to take antiretroviral therapy to prevent viral transmission to the child during that time period. Therefore, pregnant or nursing women will be excluded from science essay this study. Any woman of childbearing potential must have a negative pregnancy test within two weeks prior to study entry. All participants who engage in heterosexual intercourse must use two forms of reliable birth control for the duration of the study. When I entered the critical thinking education NIH/Oxford/Cambridge Scholars program I identified Mark Connors and Quentin Sattentau as two HIV researchers with whom I would like to train with because of their focus on HIV immunology, vaccine design, and their expertise in basic immunology techniques, flow cytometry, and microscopy. I initiated contact between the essay two individuals and they agreed to start a collaborative project with me. Thinking? After studying NAbs with Mark Connors for one summer I decided I wanted to focus on antibodies that mediate ADCC.

I outlined the science facets of thinking ADCC I wanted to investigate then received feedback from both mentors. Science? I subsequently wrote the proposal that was edited by both mentors and Mark Connors made final revisions. I meet multiple times during the sales week with Dr. Connors to discuss my research progress and to plan future experiments. We also conduct quarterly phone conferences with Dr. Sattentau, maintain a regular email correspondence, and meet at conferences. Communication will continue in the same manner while I am in Quentin Sattentaus lab.

Selection of science Sponsors and coca cola vs pepsi, Institution. Selection of Institution. I decided to join Columbias MSTP program because of its tradition of essay exemplary medical education, medical research, and grendel book, unparalleled support offered to students from essay program administrators. Because of my previous research experience with HIV I knew I wanted to continue research in cola sales, this field. I decided to join the NIH/Oxford/ Cambridge Scholars Program to complete my dissertation research because it would provide me access to a more extensive network of scientists than Columbia could. Oxford has a particularly strong network of HIV investigators and the structure of its graduate program allows for a particularly specialized education. As a graduate student in the pathology department I am allowed to select seminars and science, accelerated 1-day to 1-week courses that allow me to attain skills quickly and immediately implement them into my research. Grendel Book? In addition, the NIH/Oxford/Cambridge Scholars program provides funding for essay, students to take courses offered by grendel book, any institution giving me access to virtually any educational opportunity necessary. In addition, through NIHs Foundation for Advanced Education in the Sciences Ive had the science essay opportunity to take courses that will help me towards my goal as in infectious disease investigator such as the Vaccine Development course I completed.

The NIH/Oxford/Cambridge Scholars Program is designed so that students work with a mentor at lord of the beast, NIH and one at a university. When I joined the essay program I searched for HIV investigators throughout NIH and Oxford and identified Mark Connors and capital asset pricing, Quentin Sattentau as potential mentors because I wanted to science essay conduct research that had direct implications in vaccine development. Capital Asset Pricing Model? Quentin Sattentau was studying how to elicit effective anti-HIV neutralizing Abs (NAbs) for vaccines and essay, adjuvants to stimulate systemic and mucosal immunity. Through my search at the NIH I identified Mark Connors as a potential collaborator though he and Quentin had never collaborated before. Cola Vs Pepsi? I believed Dr. Connorss expertise in NAbs of science long-term nonprogressors (LTNPs) would complement Dr. Sattentaus vaccine development goals.

In addition, Dr. Connorss unique cohort of LTNPs would provide ready access to samples to study many facets of HIV infection. Finally, I believe Dr. Sattentaus approach to research as a PhD and Dr. Connors as an MD will teach me to think about science in a more practical way that is suited to me as a future MD/PhD researcher.

I will follow the principles on enslaved africans, responsible conduct of research as outlined in essay, NOT-OD-10- 019. Grendel Book? As an undergraduate at UMBC I took a semester-long course entitled Ethics/Integrity in Scientific Research which first help me to understand the responsibility scientists have to one another and medical community honestly share information. Science? As a Columbia medical student, I have taken the capital Hippocratic Oath to guide my patient care and I am expected to adhere to the code of ethics as outlined in student handbook. Science? Upon joining the NIH I took an online ethics course and will continue to take refresher courses annually. At Oxford I will continue my ethics training by taking the required Ethics: Introduction to Research Ethics. Since I will have access to sensitive patient data it is cola vs pepsi essential that I maintain the confidentiality of these individuals. When I started research at science essay, NIH I participated in a patient confidentiality seminar. Oxford offers an Ethics: Confidentiality course in which I will enroll to better my further my understanding and ability to enslaved africans maintain ethical practices with regards to science research.

As a physician, I hope to education alleviate an individuals immediate suffering; and as a scientist I hope to contribute to long-term solutions that can prevent suffering. To work most effectively at the interface of medicine and science essay, science, I have endeavored to earn an MD/PhD. I believe that the combined degrees greatest value lies in the perspective that it affords: to approach patient care with a researchers eye, and to approach research with a physicians focus on lord, improving the quality of life. A few years ago I conducted research on telomerase, an enzyme that maintains healthy chromosome length. When telomerase is upregulated, as it is in 90% of carcinomas, cells can propagate indefinitely. Essay? When telomerase levels are halved, a bone-marrow failure disease called Dyskeratosis Congenita (DC) results. While looking at coca, yeast cells with low levels of telomerase and measuring their growth rates, I noticed two things. The first was that cells with low levels of essay telomerase grew slowly. Coca Cola? Secondly, these cells had abnormal morphologies, as do tumor cells. Science Essay? These observations led me to wonder whether the critical thinking education progression of cancer in individuals with DC was slower than in patients without it because DC patients have less active telomerase. Upon discussing this with my P.I., he acknowledged having never considered his research from that aspect.

A literature search demonstrated a lack of science essay any substantial research concerning this question. Grendel Book? Research in biochemistry affords many new advances in medicine, but through that experience the science essay importance of keeping a broader medical picture in mind became clear to online me. My goals as a physician scientist are to essay treat patients and conduct research that will allow me to impact patients I will never meet through therapeutic discovery; specifically serving as a creative link between science and medicine in cola sales, the field of infectious disease. The training I will receive in Oxfords doctorate program will prepare me to be a competitive investigator through a tailored curriculum, expert dual-mentoring, learning to work in a collaborative environment, and by giving me extensive writing experience via paper submissions and science essay, even through this grant. These are all specific skills that would be harder and africans, take longer for me to science garner through a traditional medical student experience. The advantage I have in understanding that I want to be a physician-scientist has given me the lord of the opportunity to carefully cultivate these skills early in my career. Activities Planned Under This Award. Research 80%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10% Research Skills : While working in Mark Connorss lab I have acquired specific skills such to creatively study immunology such as learning to use flow cytometry. I discuss my research progress frequently during the week with Dr.

Connors. We also have phone conferences and essay, meet with Dr. Sattentau at numerous conferences throughout the year. Writing Skills : When I entered the program I took a grant-writing workshop sponsored by the NIH Graduate Partnership Program. I also completed a 3-day grant-writing course during my senior year of college. I will improve my writing skills through preparation of manuscripts based on my research to be submitted for publication, applying for training grants, and submission of a report for my transfer viva (qualifying exam). I will also complete the Scientific Writing from a Readers Perspective seminar. Discipline specific learning : I completed the 3-day Vaccines: Development and Evaluation of Efficacy course offered by FAES in February 2010. I also attended Psychiatry and grendel book, HIV, a 2-day workshop offered by the American Psychiatric Association so I could learn to science essay effectively interact with HIV+ patients I encounter. I attend weekly seminar series hosted at beast, the NIH such as the Wednesday Afternoon Lecture Series, Immunology Interest Group seminar series, and weekly lab meetings.

Bench to bedside clinical training : I will attend the Clinical Research Training Program (CRTP) weekly rounds. Science? I will also have the opportunity to be mentored by an infectious disease clinician. Presentations : I will continue to annually attend and present at pertinent Keystone Symposia and the Annual AIDS Vaccine conference. I have also had opportunities to present posters at the NIH/Oxford Cambridge Scholars Program Colloquium and the Oxford Pathology Graduate Student Symposium. I have given a talk to my graduate program journal club and will have more opportunities to present my research in lab meetings.

Career Development : The NIH/Oxford/Cambridge Scholars programs hosts a seminar series where leaders in government/academic research, the pharmaceutical industry, and finance speak with us about their career. I also attended NIAID Fellows retreat that focused on career development. Teaching : As I prepare to leave the lab I will be responsible for training a new research fellow. Year 1 (continued) and Year 2: Quentin Sattentau Lab (Oxford) Research 75%, Seminars/Courses 10%, Presentation/Career Development/Clinical 10%, Teaching 5% Research Skills : While working in tu clothing online, Quentin Sattentaus lab I will learn to apply microscopic imaging to studying HIV infection. I will have weekly meetings to discuss my research progress. We will have phone conferences and meet with Dr.

Connors at various conferences throughout the year. I have also been assigned a graduate advisor within the science essay pathology department but not in my specific lab who will help me to objectively evaluate my research. Writing Skills : I will improve my writing skills through preparation of manuscripts based on capital asset model, my research to be submitted for publication and writing reviews. I will complete the following 1-day courses offered at Oxford: Writing Skills Paper and science essay, Thesis; Doctoral Thesis Writing, Discipline specific learning : I will participate in courses and lord beast, Skills Workshops offered by Oxford including Advanced Light Microscopy, Electron Microscopy, and Introduction to science essay Statistics. Bench to bedside clinical training : I will attend the Nuffield Department of Clinical Medicine Grand Rounds and shadow an infectious disease clinician. Career Development : See above. I will also attend the 4-day Oxford GRADschool, a personal development course. Teaching : I will seek to be an undergraduate lab instructor and take the Oxford course Teaching and Learning Skills to coca cola vs pepsi sales prepare myself. Year 3 and Year 4: Medical School (Columbia) In the summer after 11th grade I participated in the intensive 6-week Gene Search Program at the Catholic University of America that introduced me to molecular biology and the exciting race to sequence the human genome.

That summer, I learned much more than how to essay run a gel; I learned to asset pricing model read and comprehend primary research articles and how to design experiments. Science? Because of my accomplishments that summer, I was invited to enslaved africans participate in science, an extension of the enslaved Gene Search Program during my 12th grade year. This program inspired me to pursue a career in biomedical research because I began to understand how the principles I was learning in science essay, school fostered scientific discovery. As a high school senior, I worked with Dr. William Winter at cola sales, Howard University School of science Medicine to investigate pathways that regulate 2,3-diphosphoglycerate (2,3-DPG) levels in red blood cells (RBC). 2,3-DPG lowers the affinity of hemoglobin for of the, oxygen, thereby facilitating oxygen release to tissues. Patients suffering with sickle cell anemia have high levels of essay 2,3- DPG. While this effect may seem advantageous to patients with sickle cell anemia, deoxygenating hemoglobin ultimately causes red blood cells to sickle by promoting hemoglobin polymerization.

Previous studies indicated that adenosine might regulate 2,3-DPG levels through an tu clothing, interaction with the A3 receptor. To test this idea, we indirectly measured 2,3-DPG levels by using spectrophotometry to measure a by-product of the 2,3-DPG pathway. Science? An exciting therapeutic prospect of our research is that an antagonist of the A3 receptor may prevent the grendel book accumulation of 2,3-DPG and RBC sickling in essay, patients with sickle cell anemia. I documented this work in a five-chapter thesis, in accordance with my high schools Science and Technology Program, and presented it at my schools science fair, where I was awarded third place in critical thinking, biochemistry. During college I spent four years working with Dr. Michael Summers to investigate Moloney Murine Leukemia Virus (MuLV) dimerization. Understanding dimerization is science key to enslaved africans developing gene and antiviral therapies. Essay? MuLV is similar to HIV in that it is a retrovirus that packages two copies of pricing its RNA genome.

Packaging of dimerized RNA is mediated by interactions between the nucleocapsid (NC) domain of the assembling Gag polyproteins and a specific domain of the RNA called the science essay Psi-site. Sales? Previous work in the Summers lab suggested a riboswitch mechanism for MLV genome packaging, where, in monomeric RNA, Psi-site UCUG segments capable of science binding NC are sequestered by base pairing, but upon tu clothing dimerization, become exposed to bind NC. My project was to investigate whether similar sequences in the Psi-site could also bind NC. Using isothermal titration calorimetry and nuclear magnetic resonance, we identified short RNAs (ACAG, UUUG, and UCCG) that bind NC with nanomolar affinity. Science Essay? Our findings suggest that: 1) binding depends on an unpaired guanosine, 2) binding is enhanced in short RNAs containing terminal phosphates, and 3) binding affinity varies by more than one order of magnitude depending on which three nucleotides lie upstream of the coca guanosine. The paper I co-authored (Dey, A., et. Essay? al., 2005) extends an established model for genome recognition, in which the NC domains of assembling Gag molecules interact with multiple X(i-3)-X(i-2)-X(i-1)-G(i) elements (X is a variable nucleotide) that appear to be preferentially exposed in the dimeric RNA. I presented this research at the 2004 ABRCMS and at multiple UMBC symposiums. I continued to thinking investigate potential dimerization and NC-binding regions of the Psi-site using native and mutant MuLV RNA segments in gel-shift assays. As expected from our riboswitch hypothesis, we observed NC binding preferentially to RNA dimers which was also confirmed by equilibrium dialysis. Most recently our lab published the NMR structure of this dimer (Miyazaki, Y., et al., 2010). During the summer of 2005 I participated in the Weill Cornell/Rockefeller/Sloan Kettering Gateways to the Laboratory Program.

I worked with Dr. Nikola Pavletich toward solving the crystal structure of the Saccharomyces cerevisiae Rad4-Rad23 complex bound to a DNA substrate. Rad4-Rad23 is a homolog of human XPC-HR23B, a DNA damage-repair protein. In its mutated form, XPC-HR23B causes the disease Xeroderma Pigmentosum. Science? Patients are extremely sensitive to enslaved UV-light and develop skin cancer during childhood. I identified DNA substrates to be used in essay, crystallization trials by HPLC-purifying DNA, performing gel-shift assays, and setting up crystal trays. Enslaved? I presented this work in a talk before the Tri-Institutional community and during posters sessions at essay, the 2005 Leadership Alliance National Conference and at thinking, the 2005 SACNAS National Conference. In 2006 I studied telomerase assembly in the lab of science essay Dr.

Thomas Cech at the University of Colorado at Boulder. The central role telomerase plays in protecting chromosome integrity places telomerase at the heart of two common human ailmentscancer and aging. To study how and where in the cell telomerase is assembled from its RNA and protein components, we used the budding yeast Saccharomyces cerevisiae as a model organism. We sought to understand whether the noncoding telomerase RNA (TLC1) goes through a cytoplasmic phase during its biogenesis. One model is that a polyadenylated precursor form of TLC1 is exported to the cytoplasm where it associates with Sm proteins. Mtr10p then plays a role in importing the TLC1-Sm complex back into the nucleus, where the telomerase holoenzyme is assembled. Using cellular fractionation and real-time RT-PCR, we attempted to localize TLC1 in its various stages. During the summer between my first and second year of medical school I joined the lab of Mark Connors where I screened patient sera for breadth and potency of neutralizing Abs against HIV using a TZMBL neutralization assay. We demonstrated that that progressor serum has greater breadth in potency in neutralizing HIV than Long-Term Nonprogressors (Doria- Rose, N., et al., 2009). Studying NAbs peaked my interested in other anti-viral mechanisms mediated Abs and lead to my proposed project of investigating antibody-dependent cellular cytotoxicity.

Each research experience has shown me the many nuances of a research career. I learned about: 1) the importance of basic research in developing medicines, 2) the time, organization, and ingenuity it takes to conduct research, and 3) the grendel book vital role that a physician-scientist plays in translating basic research into medical treatment. Although Ive experienced frustrations and disappointment in research, the thrill of observing something new, finishing a project, or publishing a paper always outweighs these challenges. Each successive experience only increased my eagerness to improve health care through innovative research. [Redacted from sample.] QUENTIN SATTENTAU MENTOR STATEMENT. [Redacted from sample.] [Our Web version of this sample application can't reproduce the science layout of the enslaved actual summary statement. See below for the data fields and critiques.] Center for Scientific Review Special Emphasis Panel. Fellowships: AIDS Predoctoral and Postdoctoral. RESUME AND SUMMARY OF DISCUSSION:

In this application, the science fellowship candidate proposes to tu clothing online investigate antibody-dependent cellular cytotoxicity (ADCC) in HIV-infected patients do determine the extent of protection that ADCC provides. The reviewers noted that the candidate is outstanding, with a strong record of science essay research training and accomplishment. The candidate appears to be highly motivated. Her sponsors, at both institutions, are well qualified to grendel book provide the necessary training and the letters of science recommendation were uniformly laudatory. Minor weaknesses of the application included the ambitious research plan to be accomplished in two years, the risk of conducting the research at two sites, and model, the failure to address pitfalls and alternatives of the science essay research project. These weaknesses, however, did not detract from the reviewers enthusiasm.

This training project will have a high impact in ensuring that the critical thinking education candidate maintains her potential to become an independent clinical researcher. DESCRIPTION (provided by applicant): Understanding the basis of an immune response that controls infection or provides sterilizing immunity remains a major goal in the search for effective vaccines or immunotherapies for HIV. Antibodies (Abs) induced by candidate vaccines to the surface envelope glycoprotein have not neutralized a broad array of science primary virus isolates. For this reason, eliciting a cytotoxic cellular response has been the thinking primary goal in most recent vaccine trials. Science? However, this approach has not been successful in africans, containing viral replication in vaccinees that have become HIV-infected. Antibody-dependant cellular cytotoxicity (ADCC) has been shown to mediate sterilizing immunity against challenge with pathogenic simian immunodeficiency virus [Hessel 2007]. In ADCC, Fc-bearing Abs bind viral epitopes coating an infected CD4+ target T cell and an Fc receptor bearing effector, most commonly natural killer cells (NKs), bind the Ab and use perforin to deliver granzymes which induce apoptosis in the target. We want to study ADCC in infected patients to understand the essay magnitude and characteristics of the enslaved africans best responses achieved by natural infection.

First, we will compare ADCC mediated by the sera of a cohort of patients using a granzyme B cytotoxicity assay developed in our lab. Based on these findings, we will select the sera of patients with the science essay most ADCC, generate monoclonal Abs (mAbs), and grendel book, characterize the mAbs based on science, epitope specificity, affinity, potency, breadth, IgG isotype, and of the flies, Fc type. We will also evaluate whether ADCC is disparate from classical neutralization. Finally, we will use microscopy to examine the synapse between effectors, Abs, and targets. Essay? The outcome of this research will provide insight into the characteristics of online Abs that mediate ADCC that are likely important goals in the design of essay HIV vaccines or immunotherapies. Hypothesis: Antibody-dependent cellular cytotoxicity (ADCC) is a function that has been shown to mediate protection from lentiviral infection. We hypothesize that variations in ADCC activity of enslaved africans sera are dictated by the amount, specificity, and subclass of science HIV-specific antibodies. Aim 1: Characterize the online potency of sera of HIV-infected individuals in ADCC.

Aim 2: Characterize the specificity and breadth of antibodies with ADCC activity. Aim 3: Characterize the essay structure and function of the target-effector synapse using both fixed and live cell laser scanning confocal microscopy (LSCM), transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM) and tomography. Understanding the role of antibody-dependent cellular cytotoxicity (ADCC) in tu clothing, HIV could provide important insights for induction of this activity through vaccination. This project seeks to characterize the essay Abs that mediate ADCC and image the critical thinking functional synapse formed by cellular components involved in ADCC with the essay goal of defining new goals for the development of HIV vaccines and therapeutics. Fellowship Applicant: 1. Sponsors, Collaborators, and online, Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 2. Overall Impact/Merit : This proposal is from an applicant who is currently enrolled in science, the NIH M.D./Ph.D.

Partnership Training Program and capital pricing, is a participant in the NIH/Oxford/Cambridge Scholars Program, which promotes joint research in an NIH laboratory and in a university research lab. Ultimately, she will receive her Ph.D. from Oxford University and complete her medical training at Columbia University. The applicant is highly recommended by previous research mentors and has an outstanding academic record. Her research experience, which includes four years of undergraduate research, is matched by her productivity; she is science a contributing author on two published papers and has presented her work at numerous scientific venues. Enslaved? The research plan, which derives directly from the expertise and interests of the mentor, focuses on relationships between antibody-dependent cellular cytotoxicity (ADCC) in HIV-1-infected patients, HIV-1-associated disease progression, and antibody subtype. Additional experiments, which will draw on the expertise of the co-sponsor, will investigate the target-effector synapse during ADCC. These studies will be significant for their basic science and translational value. The training plan is unique in science, that training and research will be accomplished in two separate laboratories of the enslaved africans sponsor and co-sponsor. This proposal outlines an excellent training opportunity that will greatly benefit the applicant and essay, her potential for establishing an asset pricing model, independent research career. The applicant, who is currently enrolled in an M.D./Ph.D. training program, is described in three strong letters of recommendation as enthusiastic, articulate, knowledgeable, highly focused, and very inquisitive.

Her undergraduate research mentor characterized her as easily among the top two or three of the more than 200 current and former undergraduates that he has mentored. The applicant has four years of undergraduate research experience, which involved work with the Moloney Murine Leukemia Virus, and research performed during a summer program at the Howard Hughes Medical Institute focused on studies of essay telomerase. Technical approaches for which she was trained included cell growth assays, transformation, RT-PCR, isothermal titration calorimetry, NMR, and cryo-electron tomography. Through her prior research experience, the critical thinking education applicant has been included as a contributing author on two published papers and has presented eight posters and one oral presentation at various local, national, and international meetings. Science? The applicant has an outstanding academic record (3.97 GPA) and numerous awards and recognitions reflecting superior academic performance. The collaborative project outlined in the proposal was an outgrowth of a dialog initiated by the applicant between the sponsor and co-sponsor. The applicant has provided thoughtful goals for africans, developing a career as both a scientist and physician. 2. Science Essay? Sponsors, Collaborators, and Consultants: The applicants mentor and enslaved, sponsor, who is currently Chief of the HIV-Specific Immunity Section in science essay, the Laboratory of Immunoregulation, NIAID/NIH, is an established and productive senior investigator with research support provided by critical thinking, the NIH. The co-sponsor is also a productive researcher with a consistent record of support for studies related to science HIV/AIDS.

The research interests and expertise of the sponsor and co-sponsor will provide excellent support for asset model, the applicants research and training. The expertise of the essay co-sponsor will be particularly relevant to studies outlined in Aim 3. The sponsor has trained an average of four students per year over 20 years, suggesting an excellent training environment for the applicant. Similarly, the co-sponsor has trained nine Ph.D. Grendel Book? trainees since 2003. During her predoctoral fellowship, the applicant will be interacting with four post-baccalaureate students and one M.D. in the sponsors lab, and four Ph.D. trainees in science essay, the co-sponsors laboratory. The latter trainees will overlap with the applicant for one year or more. These interactions will enrich the applicants training. The research plan is built on studies of antibody-dependent cellular cytotoxicity in lord of the flies, HIV-1-infected individuals. Experiments will examine the potency of sera of science HIV-1-infected individuals in ADCC (Aim 1), the specificity and breadth of antibodies with ADCC activity (Aim 2), and the structure/function of the target-effector synapse that mediates ADCC. The central feature of the proposed research is a novel and quantitative ADCC assay (developed in the sponsors lab) that offers significant improvements over other ADCC assays.

Results generated from experiments in Aims 1 and 2 will have both basic science and translational value, while results from Aim 3 experiments will provide basic information about the mechanisms of ADCC. The applicant and tu clothing online, sponsors are well positioned to perform these studies, since the patient populations and study procedures are already in place. The applicant is participating in the NIH/Oxford/Cambridge Scholars Program, which promotes research with a mentor at the NIH and another mentor at a university. Consistent with the Scholars Program, the research training plan includes studies and training in essay, year 1 in the sponsors lab at the NIH, continued studies and training in lord flies, year 2 in the co-sponsors lab at Oxford, and years 3 and 4 in medical school. Training in science essay, each laboratory will include the development of specific laboratory skills, writing skill development, discipline-specific education, bench-to-bedside clinical training, career development, and opportunities for presentation of results and teaching. Training at the NIH (sponsor) will be a continuation of the current laboratory work, with increased emphasis on independent research and critical thinking.

Training at Oxford (co-sponsor) will include skills development in africans, experiments involving infectious HIV-1, microscopic imaging, and biochemical and essay, serological techniques (such as ELISA, gel electrophoresis, and lord of the beast, mass spectrometry). Although the targeted patient enrollment is science essay given as 300 HIV-1-infected individuals, the experimental plan does not provide details regarding the scope of studies in Aim 1. Correlates of ADCC to be studied in lord of the flies beast, Aim 1 (rates of progression and viral loads) were not described in science essay, sufficient detail. How will rates of progression be measured? Will viral load be documented on tu clothing online, a continuous scale or will patient viral loads be stratified? An explanation of how the results of essay Aim 1 experiments will be used to grendel book guide the selection of pooled patient sera in Aim 2 would have been informative. A research project spanning the laboratories of two different principal investigators will offer diversity in training and acquired expertise, and science essay, enhance the capability of the coca sales applicant to develop an independent and productive research career. 5. Institutional Environment Commitment to Training: The resources and essay, facilities at both institutions will provide outstanding environments for training and research. A more detailed plan regarding the coordination of grendel book training plans between the science essay sponsor and co-sponsor would be valuable, considering the thinking distance and time difference between the two performance sites.

Training in the Responsible Conduct of Research: Comments on Format (Required): The description of RCR training is brief and does not provide a detailed plan for essay, formal training. Enslaved? However, enrollment in science essay, two formal courses at pricing, Oxford (both focused on ethics) is mentioned. Comments on Subject Matter (Required): Training courses and certifications at the NIH and Oxford should provide information on relevant subjects. Comments on Faculty Participation (Required): Faculty participation could not be evaluated from the description of science RCR training. Comments on Duration (Required):

The duration appears to be adequate, although exact contact hours were not provided. Comments on Frequency (Required): The frequency of online training could not be discerned from the description of RCR training. Fellowship Applicant: 2. Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2.

Training Potential: 2. Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This is an outstanding proposal from a highly qualified applicant. The applicant has an exemplary scholastic record, strong research history and outstanding letters of reference although she lacks a first author publication. The applicant has chosen two superb sponsors with complementary skill sets. In fact, the applicant brought these two investigators together herself. Science Essay? Both sponsors have excellent training track records and grendel book, both are actively involved in the HIV arena.

The primary goal of this project is to characterize antibodies that mediate antibody-dependant cellular cytotoxicity (ADCC). The Connors lab at the NIH has developed a novel flow cytometric-based assay to measure granzyme B delivered to an HIV infected cells. Using this novel assay the essay applicant plans to compare ADCC in HIV+ subjects with various rates of disease progression and to define the enslaved africans specificity and breadth of antibodies with ADCC activity (Aim 1 and 2). The third Aim to characterize the structure and function of the essay target-effector synapse will be carried out at Oxford under the flies guidance of science essay Dr. Sattentau.

This aim will use fixed and live cell laser scanning confocal microscopy and TEM to examine the samples between NK and other cells with ADCC activity. If the applicant is successful the data from this project will be highly important for critical thinking education, HIV vaccine development. The research project however is essay quite ambitious and no timeline is given. It appears that the applicant plan to enslaved spend 1 more year in the Dr. Connors lab and 1 year in Dr. Essay? Sattentau lab making it a very busy two years if the applicant plans to finish all the work they outlined. Despite the highly ambitious nature of the proposed research project the overall project goals warrant further study and the applicant herself is of the highest caliber making this application worthy of funding. The applicant has a very strong research background dating back to high school where she participated in the Gene Search Project at Howard University Hospital. As an undergraduate at UMBC she was a member of the Meyerhoff Scholars Program funded by the HHMI. The applicant has coauthored 2 papers and is the sales first author on 8 posters and 1 oral presentation. The applicant has extremely strong letters of recommendations, one of which is from science essay a Nobel Prize winner.

Exemplary scholastic record and lord, many community-based volunteer positions. 2. Sponsors, Collaborators, and Consultants: Two suburb sponsors are listed. Dr. Connors, Chief, HIV-specific Immunity Section NIAID, has a long track record of science essay high quality research and publications and a solid list of lord beast successful trainees (4/year for 20 years). The other sponsor, Dr. Sattentau, a Professor of Immunology and Pathology at the University of science essay Oxford, is equally meritorious with over 151 published manuscripts and solid funding record. Dr. Sattentau, also has a established track record of training PhD student (n=9) and post-docs Both sponsors have well established complementary HIV research programs that will support the asset model applicants proposed research project. The planned research, if successful, could provide important information about of the role of ADCC in HIV infection.

The project is well outlined and there are sufficient preliminary data to support the feasibility of the project. The plan is to utilize a novel recently developed (Connors lab) quantitative flow cytometric assay to measure GrB cytotoxicity and determine which Abs are capable of mediating the essay highest level of ADCC. Using this technique the applicant plans to characterize the potency of sera from subjects with different rates of disease progression, characterize the specificity and breadth of capital asset pricing antibodies with ADCC activity. Using advance microcopy techniques (fixed and live cell laser scanning confocal microcopy and electron microscopy) the applicant plans to examine the structure and function of the essay target-effector synapse. This work will be done under the guidance of Dr. Sattentau who is an tu clothing, expert in studying the virologic synapses. This is a highly ambitious grant. It may be difficult for the applicant to finish all the proposed work especially the work proposed in Aim 3. The applicant is calling for 300 subjects but no power calculation is shown to science essay support that many subjects. A timeline for the proposed work would be helpful. The applicant does not anticipate any issues nor are any alternative approaches outlined. This maybe a little naive.

The research training plan is well laid out by year. Tu Clothing Online? The plan includes research, writing skill (grant writing course), presentation, career development, teaching (both in class room training of new personnel) and discipline specific learning (i.e. in house courses and seminar series, lab meetings). Letters describing each institutions role in training the science applicant were provided. Applicant states that she will interact on daily basis with section head and other senior laboratory member on enslaved africans, a daily basis. Students also make formal presentations and attend weekly LIR, meeting, NIAID grand rounds, clinical center Grand Rounds and HIV journal club. The training plan indicates that the applicant will be at the NIH for one year and Oxford for one year and then back to medical school for the last 2 years. This raises some concerns regarding the essay duration of the funding (3 vs. Capital Pricing Model? 2 years). 5. Institutional Environment Commitment to science Training:

The work will be conducted at the NIH/NIAID and University of model Oxford. Both institutions are of the science essay highest caliber with access to all the required resources. Oxford has a program in place for graduate students to take accelerated courses and the NIH Foundation for Advanced Education in the Sciences also has specialized courses that will facility the applicants goal to grendel book become an infectious disease investigator. Training in the Responsible Conduct of Research: Comments on Format (Required): On line ethics course at the NIH and formal classroom course at Oxford. Comments on science essay, Subject Matter (Required):

Applicant states they will follow principle of research outline in NOT-OD-1-019. The applicant has taken semester long course on Ethics/Integrity in Scientific research. The applicant took another Ethic course upon entering the NIH. Oxford offers a similar course. Comments on Faculty Participation (Required): No comment on vs pepsi sales, faculty participation. Comments on Duration (Required): Comments on essay, Frequency (Required): Online course and annual refresher at of the beast, the NIH. Additional Comments to Applicant (Optional): Fellowship Applicant: 1.

Sponsors, Collaborators, and Consultants: 1. Research Training Plan: 2. Training Potential: 1. Institutional Environment Commitment to Training: 1. Overall Impact/Merit : This proposal comes from an excellent candidate who is part of the NIH/Oxford/Cambridge Scholars program, carrying out her PhD research under the guidance of two excellent researchers and mentors, Drs.

Connors (NIH) and Satentau (Oxford) and will complete her MD studies at Columbia. The primary hypothesis of this proposal is that ADCC activity levels in sera are dictated by the amount, specificity, and subclass of science essay HIV-specific Abs. Tu Clothing? This represents a highly relevant area of research that will be tackled through 3 straightforward and well-planned aims (i) characterize the potency of sera from individuals at different stages of HIV infection in ADCC (comparing to clinical status and Ab neutralization), (ii) characterize the specificity and breadth of Abs mediating ADCC (comparing the sera to panels of known neutralizing Abs), and (iii) characterize the ADCC synapse. This will involve state-of-art approaches established in the sponsors labs, allowing the applicant an excellent chance to perform this research. Science? It is a clear and well-written proposal detailing a strategy to carry out relevant research in an excellent training environment.


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Song of Solomon by Toni Morrison essay. Song of Solomon is a social commentary on racism and its negative effects in the society, abandonment and a journey of self discovery. Morrison, through the science, journey that Milkman undertakes, is able to lord of the flies, make her audience examine racial relations in the American society. Song of Solomon is a tale of racial injustices and an epic journey by Milkman to find his roots and ends up finding himself. Milkman is a selfish African American male brought up in a life of privilege and oblivious of racisms that engulfs his community. He goes through most of science his life without even thinking about the suffering of his community and suffering he causes to those who relates to. Until a quest to thinking education, find gold that his father and essay, sister once saw in cave turns into a self discovery journey. His journey takes him through the suffering of of the his parents and grandparents as well as those experienced by his community. Songs of Solomon is a statement on powers of science traditional songs and stories and its power to enslaved africans, preserve history, especially where written form does not exist. Folklore, traditional songs and stories were used by the African America to preserve and science essay, pass on their rich history from one generation to the next. Critical Thinking. Milkman is able to trace his family history from stories and science essay, songs and in the process he discovers his roots and gets reattached to his community.

Morrison uses Milkman’s journey to communicate the power of stories, folk songs and folklore as they become a compass that helps Milkman find way in his epic journey. Songs of Solomon is also a social commentary against racism in thinking the 1960s, in America, and women oppression. Essay. She takes her audience to Milkman’s journey of self discovery and uses an old folksong to capital, aid him in tracing his past. Morrison gives her audience a unique look into the social issues of the American society in science essay early years of the slave emancipation and their illusion of freedom all the way to enslaved, the 1960s. Science. She uses flight as a way for people to escape their suffering and their problems.

The story begins with an attempt of an capital model insurance sales man to science essay, fly from a hospital’s rooftop. His audience in the ground comprising of socially stratified African Americans looks on without trying to stop him. Most of people in the watching crowd reject the belief from traditional stories, songs and folklore that it is possible for an individual to fly to escape from cola suffering. His flight turned him into a folks’ hero from one generation to the next even though he had abandoned his wife and twenty one children. His escape from slavery was admired while putting his family into great suffering.

Flight is one of the strongest themes in Morrison’s book. It is depicted as a means of science essay escaping constricting circumstance in the life. Solomon Milkman’s great grandfather had flown to Africa to escape from slavery. His story of escape was passed on from one generation to the next in the folksongs and capital model, stories. To his community, Solomon is a hero; although, he abandoned his large family of science essay twenty one children and tu clothing online, left it in the hands of his weak wife. On the other hand, she is unable to essay, come to capital model, terms with the abandonment and dies in despair. In her community, her escape from her predicament through death is seen as a sign of weakness. Milkman’s escape from his desperate directionless privilege life is also perceived as a flight from the desperate circumstances (White).

His flight turns into science, a blessing in disguise for a man who at cola vs pepsi, one point is unable to figure out who he is. In his quest to find gold, Milkman finds something more precious than this metal; himself and his true name. Macon Milkman Dead III, as his name suggests, is able then to escape from the “dead” or his dead existence through flight. He is also able to figure out essay that his aunt Pilate is able to lord, fly without leaving the science, ground. She is able to do this through her love of live expressed through her helpful attitudes toward those surrounding her. Enslaved Africans. Additionally, she is able to live her life without relying on anyone, despite having a rich brother. This makes her one of the strongest characters in the novel.

She does not let her appearance divine her or the perceptions of those surrounding her rule her life. Milkman, on the other hand, is science, trapped by how the society perceives him. He walks with a strut to hide his short leg so as the society cannot see his flaws and instead he gets more alienated. His struggle to get accepted by his society blinds him from perceiving the reality they live in. Abandonment is the grendel book, other strong theme in this novel. The heroic Solomon takes his flight and essay, leaves behind his family to suffer while he gains his freedom. This causes his wife’s death as she cannot come to terms with his abandonment.

This implies that to gain his freedom Solomon has to abandon his large family. His wife through death abandons her children and coca cola vs pepsi sales, suffering. Milkman has to abandon his life of privileges to discovery himself. This means to leave behind all his comforts in order to become free. Science. Morrison is able to show the double suffering through abandonment that women in African American communities were exposed to. Solomon’s wife, Ryan, was abandoned by her husband as he escaped through his flight to Africa to drop the bondages of slavery. His wife, however, has been left not only a slave but with a large family to feed and when she dies out of despair, the society perceives it as failure while Solomon is revered (White). Guitars’ mother undergoes the same kind of predicament, she is left ill with kids to feed and care for, when her husband dies, she is therefore left without a helper.

Milkman’s leaves his adoring girlfriend to suffer when he embarks on cola sales, his self discovery journey, she eventually dies because of his rejection. In this novel, racism is the cause of most of the suffering and problems, experienced in this society. It is, therefore, another strong theme in the novel. In this book, racism changes the essay, lives of characters in different ways. Solomon, for instance, is bound by the bondages of slavery and enslaved africans, racism.

The African American race is perceived to be less equal in science essay the society and can, therefore, be owned by critical education their fellow men and put to backbreaking chores. Essay. After being freed by Lincoln in 1963, slaves were disillusioned by their new status as free human beings. They were not immediately accepted as equal despite the spirit of emancipation of lord flies slaves being that all human were equal. Today there are those who are yet to accept that all men are equal. Macon Dead II and his sister Pilate experienced the evils of slavery at a very early age; that of their father’s murder. This event turns Milkman’s father into a shell of man whose only pursuit in life is wealth to escape from science racism.

He sees wealth as the only factor that would make him equal to the white members of the society. He, therefore, mistreats his fellow African American as he pursues riches and passes on his uncaring quality to his son. Therefore, we can argue that in his bid to escape racism his relationship with his son is turned into a shallow father- son relationship. This implies that they are unable to enjoy the father son activity that brings joy and humanity in the society. Enslaved Africans. All he wants to teach his son is how to make as much money as possible. Indeed, he is the one who advised him to steal from his sister thinking that she hoarded the gold that they had found in the cave. Racism thus blinds him into perceiving life as a pursuit of material wealth regardless of who is science, left suffering in such pursuits.

Guitar, Milkman’s best friend, turns into a killer because of racism (White). He has no remorse committing murder, as long as he is killing the grendel book, whites for their mistreatment of the black community. He joins a revenge group that avenges every perceived murder of African American exactly on the same day, a week later with the killing of white members of the society (Morrison). Science Essay. Therefore, this alienates him not only from his community, but also the enslaved, wider society, for humanity is taken away from him. Analysis of characters and their relationships. Milkman |Dead is the protagonist in the book. He is depicted as a selfish immature man whose life of privilege has sheltered him from the realities of life of an African American in his era. He is oblivious of the equality and the struggle of his community. He treats those who love him with disdain and it is essay, oblivious due to the fact that racism is asset, a reality within his community.

Although Morrison treated his character as a shallow self- obsessed character in the first chapters of her novel, she nevertheless painted him as a man who cared little about anyone’s skin color. Essay. Thus, his obvious qualities make him a complex character and set the stage for his self discover journey in which he emerges as a better man. From the onset of her book, Morrison paints Milkman as a man who is always looking behind (White). He urinates on his sister as a child when he is distracted by the thought of someone being behind him. She, therefore, sets the online, stage for his journey into his family history in bid to discover himself. In other words, Milkman has to essay, journey back into his ancestors past in order to emerge a free man who truly understands his position in coca sales the world and learns the true meaning of his life and in essay the process he makes himself better. She also paints him as a very lucky character surrounded by people whose love he does not re-quite, yet they continue to love him despite his flaws. In a way Morrison is capital asset pricing, telling us that despite his shallow character those around him can unconsciously see a man with more character that he is science, aware of, but lacks purpose in his life (Morrison). She, therefore, sets the stage of his transformation early in her book. Milkman is also an outcast in of the flies his community; he has very visible flaws such as his unequal legs. His privileged up bring also sets him apart from the mostly poor African Americans in his community.

In addition, his father is an intensely disliked ruthless landlord. So despite having a rich background amidst a poor community, Milkman is more alienated for their wealth. He, however, perceives himself as a better person than those around him. His father’s wealth thus turns Milkman into a selfish arrogant man who is oblivious of the fact that most of his neighbors have to struggle for their daily necessities. Thus, his privileged background turns him into a man unaware of his environment. Milkman’s selfish nature is also evident in the way he treats his friend, Guitar. While he can easily unburden his troubling thoughts and emotions to his friend, he is essay, incapable of returning this favor. Their relationship, however, reveals a positive quality in cola Milkman. Though privileged, he is still capable of relating with a person from a lower social background. But their relationship can be argued to be a one way street, where Guitar does most of the giving.

Morrison is science essay, able to help her audience perceive Milkman’s selfishness through the eyes of his friend. In the novel, Pilate is depicted as a strong willed poor woman. Grendel Book. She is the sister to milkman’s father Macon Dead II. But unlike her brother she cares less about material possession and treats everyone with love and science, care. In the songs of Solomon Pilate represents the truly independent women. She does not let her appearance define her; she is the wine maker whose vacation in wine making enables her to survive without relying on men. Asset Pricing. This is in contrast to the other female characters in essay Morrison’s book. Thinking. In this novel most female characters are casted as helpless creatures that have to rely on men for their survival.

They fall helpless in science essay love with men who refuse to return their unrequited love. Tu Clothing Online. Pilate seems to have escaped this trap. Morrison also painted Pilate as the moral compass in essay the songs of Solomon. She is critical thinking education, a woman who is beyond reproach; she treats all people as equals and is incapable of doing any evil. Yet, there are times when she had to result into what would seem as cruel acts to stand up for others. For instance, she used a voodoo ragdoll to science, scare her brother into allowing Ruth to give birth to Milkman instead of grendel book undergoing an abortion (Morrison). This is in contrast to her evil incapable character that Morrison wants us to assume.

In that she is not afraid to scare others through mystical beliefs for someone else’s benefit. Science Essay. She also defends her daughter against an abuser by critical education scaring him with a knife. But these acts depict her as the person who is capable of science essay hurting others. Tu Clothing Online. These facts bring out her strength. This implies that she is capable of being a bit ruthless in order to science, protect others.

Her strength thus, is implied by her commitment in standing up for the seemingly weak in her community. She also stands out from most women in this community. Critical Education. Her unkempt appearance defies the traditional convections, which seem to dictate that women should be obsessed with their appearances, in order to be admired. Her own brother treats her with contempt, out of science essay her defiance of such widely held values at the time. Other members of their community treat her as if she is crazy for her refusal to conform to their perception of the values that women should represent. Capital Asset Model. Yet, she appears more comfortable with herself despite what others think of her. These qualities are responsible for strength and the final revelation to science essay, Milkman that Pilate was flying even when on ground. Her relationship with Milkman begins when he was young, even though he was prohibited by his father to visit her. He falls in love with her daughter, Hagar, and later mistreats her after using her to gratify his sexual needs. He steals gold from her as his father has told him and it is his first reason to start his journey. After undertaking his journey, he returns a transformed man.

Milkman’s relationship with his aunt turns into one of admiration. He is able to see her true nature; an independent, generous, and moral woman. Morrison’s depiction of this character is that of an capital immoral man who uses his racial hatred to justify his actions. Guitar hatred for essay white people begins at an early age with his father’s death. He holds the racism against the African American community responsible for enslaved africans his family’s predicament. He is, therefore, able to commit murders as a member of the seven days without remorse. Guitar is portrayed as deeply disturbed character in the novel, he is not even afraid to kill his friend, Milkman for science essay his perceived betray. His relationship with Milkman is complicated. Tu Clothing Online. For instance, they two are completely opposite of each other. While Milkman is depicted as a selfish individual obvious of predicaments of the African American community in their time, Guitar is completely aware of his standing in the society as a black man (Price).

He goes out of his way to punish the whites for the perceived racial injustices committed against the blacks. Morrison used Guitar to bring out the essay, indifference that Milkman portrayed towards racism. He perceives himself as a slave while his friend’s perception is thinking, that of a free man with a slave name. He is also responsible for Milkman’s seemingly symbolic transformation by trying to science, murder him. The author is, therefore, able to infer the death of the critical thinking, old Milkman when Guitar tries to choke the life out of his friend. His secretive activities as a member of the vigilant group do little to help the racial injustices in the society. Although, in the novel his acts are justified by author as those of essay a man frustrated by flies the wrong treatment of his race, he actually ends up looking more racist that those he ought to stop. In the end, Guitar fulfills the science essay, foresight of his friend, Milkman, who had foreseen him to turn into nothing more than a killer for his lack of remorse and thus murdering even the black people. His attempt to kill Milkman is, therefore, a fulfillment of his friend’s foresight.

His relationship with Milkman also helps the author portray the perception and contempt in grendel book which the authority was held by the African Americans at science, that time. Milkman had never had a running with the police until they were stopped and searched when they had stolen gold from Pilate. Pricing. Yet, Milkman was well aware of the murders that his Friend and his vigilant groups were committing against the whites. Essay. But he never felt compelled to report these crimes to the police, nor did he feel of withholding such knowledge. Through the criminal acts of Guitar and his fellow group members and the reaction of those who knew of their activities, Morrison is coca vs pepsi sales, able to communicate the relationship between the African Americans and police. To them, they were tools of oppression and did not deserve their help just as they provided none to essay, the community. Having information that could aid them resolve crimes, therefore, did not invoke any feelings of grendel book responsibility. Macon Dead II was a ruthless man who placed material wealth above anything else in his life. To him money was the answer to all the troubles that he underwent as a child and an answer to being perceived as a lesser man in the society. Macon Dead II was brought up in a family that practiced sharecropping. In his childhood, he witnessed the brutal murder of his father by white men.

Morrison infers that he saw poverty as the cause of his father’s death and racism. This is why he decided to peruse material wealth at all cost. Science. In his pursuit of material wealth Macon Dead II lost all his humanity thus, mistreated his family. He failed his son to teach how to enjoy life. Critical. Hence despite all their wealth, his family was unhappy, especially his son. Morrison was able to make her audience examine racism and its negative effects in the society through her book. The book has also a strong abandonment theme and is a tale of science essay a journey of self discovery. Morrison manages to make her audience examine racial relations in the American society showing the flies, journey undertaken by her main hero, Milkman.

Song of Solomon is a tale of racial injustices and an epic journey by science essay Milkman to find his roots and in the process ends up finding himself. Milkman emerges a better man through his quest to find his roots.

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Essay on the Types of science essay, Adaptation in Living Beings. Lord Of The Flies? The term adaptation is science sometimes used to flies, describe short-term changes which are actually a form of responsiveness. We often say that man is very adaptable to climatic conditions, for instance, since he can adjust from a warm climate to a very cold one. A more clear-cut example of responsive adaptation is science essay seen in the case of less complex organisms, such as protozoa, where certain chemical substances may be added to their aqueous environment by degrees until concentrations are reached that would have killed them is such a quantity of chemical were added all at enslaved one time. Evidently, such adjustment is possible through chemical and physical chances in the protoplasm, whose capacity for such adjustment is, of course, limited. However, the very occurrence of essay, such adjustments illustrated the tremendous versatility, of protoplasm in responding to environmental changes, and as a type of responsiveness; it is of considerable biological significance. All organisms exhibit the capacity for at least some versatility in this regard. However, this is not what is meant by adaptation.

This term describes the characteristic of living forms to develop, over a period of time, certain structural and functional features which enable them to survive and reproduce within the limits of a particular environment. Lord Of The Flies Beast? Hence, when a biologist says that adaptation is a characteristic of living forms, he has in mind a process, and one that has gone on throughout past ages, the science essay, result of which is the variety of present day organisms. However, any structural or functional feature of an organism that has been developed by this process may be called an adaptation, and so the same term is at once used to describe an overall process in nature and any given result of that process. The leaves of grendel book, cactus are greatly modified as spines, and these have very little surface area from which water might evaporate. Science? Actually, they are of little use to lord flies beast, the plant except that they prevent desert animals from eating it for the water that it contains. Science? Reduction of exposed surface and loss of photosynthetic ability by these modified leaves have been accompanied in the overall modification of the plant by an emphasis on the stem as a photosynthetic organ. Capital Model? These structural adaptations relate the science essay, plant to its environment, but they developed over time by the overall process. In general, any characteristics exhibited by an organism which re of benefit to it in critical thinking education, relation to a giver, environment are primarily either structural or functional in nature. Structural adaptations are the more obvious, and it would be difficult for science, even an amateur biologist to miss the tu clothing online, direct connection between the morphological features of almost any organism and its environment. We have already called attention to the spines and stems of cacti; these plants exhibit many other structures which make it possible for. Science Essay? For purposes of discussion, therefore, we shall distinguish between environmental factors that are physical in nature, those that are biogeochemical, and those that are biotic.

One of the most important environmental factors to which organisms are subjected is light. This should be obvious in the case of green plants since they are dependent upon sunlight as an energy source in the photosynthetic process. The leaves of higher plants are adapted in enslaved africans, various ways to sunlight; in many cases, they are arranged on the plant in such a way that maximum exposure to sunlight is realized. However, the influence of science essay, light is not limited to photosynthetic effects; the africans, flowering process in many plants is rigidly controlled by the time pattern and quality of light; received. One example such control is science photoperiodicity, to the response of an organism to the length of time it is exposed to light. The principle of photoperiodicity is exploited by commercial nurserymen in producing blooms out of season. Africans? By supplying artificial light or by subjecting plants to periods of essay, darkness, as the case may be, it is possible to regulate the time of flowering. Some species are termed long-day plants, because they normally bloom only when days are long, whereas others are called short-day plants, since they respond in their flowering processes to shorter periods of light.

As might be expected, there are species which are intermediate in their light requirements for the development of flowers, and there are some whose flowering process if not influenced at critical education all by length of light exposure. In addition to the regulation of lowering, a great many other light-controlled processes are known in essay, plants, some of which are seed germination, leaf fall, the development of tu clothing online, colour in leaves, and growth rates of plant organs. A number of adaptations to light are also seen in the animal kingdom, among the most obvious of which are eyes of higher animals. Essay? Comparative studies of animal vision reveal that a direct relationship exists between visual adaptations and environment. This, in turn, is reflected by the adaptations of flowers and berries.

Most insect-pollinated flowers are not red, whereas many which depend upon birds for pollination are. Furthermore, the berries and fruits of flies, many plants are red; as a consequence of which birds are attracted to them. Such species may become widely distributed, since birds ingest and science later eliminate many seeds. As a matter of fact, the seeds of cola vs pepsi, some plants do not germinate readily unless they have first passed through the gastrointestinal tract of an animal! This illustrated how far-reaching one factor of adaptation may be. Again, the eyes of science, nocturnal animals such as cats and beast alligators have extremely sensitive light receptors. These receptors are protected from science essay daylight intensities by grendel book slit pupils, which admit far less light than round pupils. In addition, important aspects of animal behaviour such as mass movements, reproductive activity and feeding habits are influenced tremendously by light. 'Another important factor of the physical environment is temperature. If a survey is made of essay, plans and animals that exist at capital pricing model various temperature zones of the earth, it is science essay found that there is a direct correlation between this factor and those features which adapt organisms to their respective habitats. Many animals are adapted to critical education, seasonal changes, and in some cases even change their camouflage patterns.

In addition to its influence upon animal and plant distribution, temperature is of considerable ecological significance in other ways. Seeds of most plants do not germinate until the science essay, temperature if fairly warm; some will not germinate even at the proper temperature unless they have previously been exposed to a period of cold. One rather unusual adaptation to temperature is seen in the case of critical education, seed cones of the jack-pine, which do not open readily to release the seeds until they have been scorched by fire. Apparently, this is an adaptation that enables this species to survive forest fires. Among animals, temperature is an important factor m reproduction, rate of embryonic development, migratory activities, and a great many behavioural characteristics. As an science essay, example of temperature adaptations in animals, the birds and mammals are able to maintain constant body temperatures by virtue of a feature of the nervous system which is absent in cold-blooded animals. This is the temperature-regulating centre of the brain, a kind of coca cola vs pepsi, built-in thermostat. This feature, plus accompanying heat- regulating adaptations such as the feathers and hair exhibited by these two groups of animals, respectively, makes it possible for them to exist under a wide range of temperature conditions. Water is another factor that is ecologically important.

Some organism possesses adaptations that make it possible for them to live entirely within an aqueous environment, whereas others are adapted to land. Structural adaptations in science essay, these respective environments are frequently quite obvious; gills of fishes and lungs of land vertebrate are among those that are better known. We have mentioned the enslaved africans, contractile vacuoles featured by many unicellular freshwater organisms which are adaptations to osmotic pressure. In oceans, it has been observed that certain species are characteristic of various depth zones, indicating adaptations for withstanding pressured that exist at different levels of the science essay, sea. Flies Beast? Organisms that live on land and are adapted variously to water some plants and animals are restricted to moist environments, while others inhabit only dry areas. Science? Cacti are typical plants that possess features making possible their existence in desert areas, and among animals, the camel and sales the kangaroo rat have become almost proverbial for their capacity to withstand dry conditions. In addition to these physical factors, a number of science essay, others assume great importance. Among these are chemical and nutritional substances or materials, various gases, radiations, and so on.

In general, however, light, temperature, and coca cola water may be considered the most important physical factors of the environment. It should be pointed out science that these environmental factors are merely separate parts of a total situation, and they frequently exert effects together. For instance, it was mentioned that seeds of a given species will not germinate below a certain temperature. However, even if an ideal temperature is provided, there will be germination unless enough moisture is present to activate enzymes and to critical education, enter into essay, reactions within cells of the embryonic plant. In this case, neither temperature nor water are independent of each other; a seed if adapted to germinate only when certain conditions of both factors are met. Similarly, the temperature of an environment may be ideal for some animal to exist successfully, but if its water needs are not met, it cannot survive.

This important principle is thinking education called the law of the minimum, and stated more precisely, it holds that regardless of how satisfactory one or more requirements of an organism may be, it cannot survive unless all requirements are met. As further example of this law and its application, let us suppose that a certain plant is supplied with all physical and science chemical requirements in adequate amounts except one essential element. Regardless of how adequate other environmental conditions maybe, the minimum requirements of the plant have not been met, and tu clothing online it cannot survive. Science? Put in terms of an analogy, a chain is no stronger than its weakest link. If the list of requirements for a given organism be considered a chain, and critical thinking education if only one item is missing, then the rest of the chain is worthless in terms of keeping the organism alive. Since plant and animal bodies are composed of matter, they depend upon their surroundings for those elements which make them up and keep them alive.

Significantly, matter is not ordinarily lost to the world of science, life with usage; a given atom of carbon. Since the atoms of such elements may be used over and over, we say that they travel through cycles. Because certain phases of a given elemental cycle may involve other factors than purely biological ones, we refer to the entire Pathway as a biogeochemical cycle, a term which tells us that biological, geological and lord of the flies beast chemical factors are all involved. In order to see how elements become involved in cyclic changes, let us consider two of the elements which figure prominently in living systems, carbon and nitrogen. Now let us consider the element nitrogen, which is found in quantity within all plant and animal bodies.

It is not present as molecular nitrogen, of science essay, course, which is a gas; rather, it is an essential part of proteins and online certain other types of science essay, organic molecules. Of course, just as carbon may be trapped in organic molecules for long periods of time, nitrogen may become temporarily unavailable at some point of the cycle. At any rate, the grendel book, nitrogen of waste products and dead bodies eventually appears in the form of ammonia. Further bacterial action makes possible the science essay, formation of nitrites and then nitrates. In the latter form, nitrogen becomes available to green plants, which combine it with photosynthetic products in the synthesis of enslaved africans, plant proteins.

It should be noted that a lesser cycle occurs between nitrates and atmospheric nitrogen; nitrates may be decomposed by essay certain bacteria in soil and water to asset, release gaseous nitrogen to the atmosphere, which is something of a loss to the main cycle. However, this loss is partially compensated for by the phenomenon that lightning converts gaseous nitrogen to nitric acid which becomes deposited in the soil, and by the ability of science, certain bacteria and algae to fix gaseous nitrogen in the form of organic compounds. Carbon and grendel book nitrogen are only two of the science essay, many elements that undergo cycles of this sort, and grendel book their changes in form may be considered typical of others. Because these cycles function as they do, there is made possible a constant refuse of matter. Science Essay? In the sense, therefore, matter is conserved; energy may be lost, organisms die, and species undergo changes, but the fundamental units of cola vs pepsi, matter travel their cycles in unending fashion.

A given organism may be influenced by two other groups of science essay, organisms: those that are members of its own species and those that are not. There is at least one essential relationship which much exists among members of most sexually reproducing animals. A great many adaptations are seen in both the grendel book, animal and plant kingdoms which allow for biparental reproduction. Some of these are highly specialized. With regard to essential requirements such as food and sunlight, members of the same species are frequently found to be in competition with one another.

One of the most fundamental of all ecological principles is that the reproductive potential of a species tends to run far ahead of its food supply or available space, and it is inevitable that some members survive at essay the expense of lord beast, others. As we shall see, this principle plays very important part in the establishment of new adaptations in species. Some species are so organized that cooperation, rather than individual competition, is exhibited. For instance, wolves sometimes form packs that can successfully attack animals too large to be captured by any one individual. This type of cooperative group is called an aggregation. A much more highly organized cooperative unit is the society, which achieves its higher degree of specialization in certain insect species.

In the consideration of relationships that exist between members of different species, competition is science of great ecological important. Plants of different species compete in nature for sunlight and tu clothing root space; animals whose food or shelter requirements are similar may be in keen competition, especially when food or space becomes scarce. Perhaps the most important ecological consideration of inter-specific relationships, however, is the principle of the food chain. Essentially, a food chain is a series that begins with a chemical medium, either soil or water, within or upon which photosynthetic producers serve as food for science, varying numbers of animal consumers. Critical Education? These support certain microscopic animals which, along with the essay, algae, constitute a mass of small, living forms collectively known as plankton.

Certain fishes, especially small forms, are adapted to live upon plankton by asset virtue of mechanisms which strain these organisms out of the water as it passes into the mouth over the gills. Eventually these fishes are eaten by larger forms which may in turn be eaten by others, and so on. Hence, there is science a chain beginning with the ultimate consumers. Eventually, of course, the consumers themselves die, and they may figure prominently in the same food chain or in a different one. Somewhere along the line, animal materials are decomposed by bacteria or fungi, and model the products of decomposition are made available to the producers. In fact, materials of dying and dead organisms and their wastes are decomposed by bacteria and fungi throughout the chain. Food chains necessarily involve predation, or the feeding of certain forms on others. Except for a few species of plants such as the Venus fly-trap, which we have mentioned, predators are all animals. Two classes of predatory animals are generally recognized' carnivores are adapted to feeding on other animals and herbivores to feeding on plants.

Some species exist on a mixed diet, however, and thus a third group, the omnivores, maybe recognized. Predation is frequently thought of as a somewhat unpleasant side of nature, and many persons build up some rather anthropomorphic views with regard to essay, carnivorous animals. Hence, a lion is often pictured as a bad animal, and a sheep is critical education evaluated as a good one. Some people actually think that a carnivore is capable of making value judgments, and ought not to kill and devour other animals since this is cruel and wicked. What these people fail to see is that an animal which is adapted through heredity for a carnivorous existence cannot be expected to shake off these structural and functional limitations and start eating grass. Their behaviour,, at least with respect to the acquisition of food, is quite determined. Furthermore, predation is science essay not as bizarre a phenomenon as some people might think. In the first place, many plants and small animal forms, which apparently lack any sort of grendel book, consciousness, are involved.

They are no worse off for having been eaten than a rock is for essay, having been broken. As for higher animals, there is vs pepsi no reason to believe that they are capable of the mental anguish with which human beings regard death. Although this certainly does not justify a cruel attitude, it does indicate that human values cannot be expected to apply to other animals. Aside from such considerations as these, predation is science essay very important in cola sales, the maintenance of natural populations. We mentioned that reproductive potential in organisms runs far ahead of available food and space; were it not for predators, animals and plants would overproduce, and certain forms would become extremely numerous. Such an imbalance is often seen when normal population rations of essay, nature are upset in some way.

When it is excessively hunted by man so that numbers of individuals are greatly reduced in a given area, the grendel book, result is usually one that is science ecologically worse for man than the former situation. With the decline of the coyote population, smaller animals upon which they would normally prey increase in number until they become more detrimental than coyotes. The natural prey of capital asset model, coyotes is the rabbit, whose reproductive potential is proverbial. Within a short time after the disappearance of coyotes in a given area, rabbits generally increase to such an extent that they consume pasture grass and other plants that support livestock. The rancher may thus be choosing the worse of two evils when he destroys coyotes.

To cite another example, hunters sometimes urge the eradication of wolves and mountain lions because they prey upon deer. Studies have shown quite clearly that, in the long run, predators exert a favourable influence upon the long-range survival of essay, deer populations. For the most dispensable members of the herd. Almost without exception, the elimination of predators has led to difficulties in maintaining healthy herds of of the flies beast, deer. Organisms of different species often become closely associated in science, their environmental; adaptations and critical these relationships are of special interest to the ecologist. One such relationship is essay that of mutualism, wherein two associated organisms belonging to different species both derive benefit from living together. We have observed that lichen is coca cola vs pepsi composed of an science essay, alga and a fungus, both of flies, which are adapted to live under conditions where neither could exist separately. The alga is able to furnish photosynthetic materials to the fungus, while the fungus provides a suitable environment for the alga by science supporting its cells and by holding water which it can use. We also mentioned the enslaved, association of cellulose-digesting protozoa with termites, an arrangement that enables these insects to essay, subsist on wood. At the same time, the protozoa are provided with a place to live, and they also receive essential materials such as water and inorganic salts from their mutuality partners. Another type of relationship is that of commensalism, in which two members of different species are so associated that one derives benefit while the other is neither harmed nor helped.

By virtue of this arrangement, a given barnacle may take in bit of food that drift away from the grendel book, other organism when it eats, and in terms of species distribution, it may be carried to points it could not possibly reach otherwise. At the same time, it does not harm to its partner organism. A third important special relationship between members of different species is that of parasitism in which one member. Essay? Parasites such as these that live within the tu clothing online, body of the host are known as end parasites. Others, called ectoparasites, exist upon the bodies of animals or plants. Science? Lice, ticks, and leeches are representative ectoparasites. Preserve Articles is home of thousands of articles published and preserved by users like you.

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ceo essay roadmap I remember when I first fell in love with cars. It started small with Hot Wheels when I was three and Micromachines when I was six. Everything about them was fast and exciting even the commercials were narrated by science essay the Worlds Fastest Talker. I loved them. Then, when I turned 12, my dad and I began taking annual trips to see the real thing at the New York International Auto Show. I looked forward to going every year, because even at coca cola sales that young age, I felt a connection to cars and the freedom they represented. I think, in some ways, it was my love of cars that largely influenced how I saw the world.

But it wasnt until I took a life-changing city planning course in college that I had an epiphany: Cars werent just shaping my worldview; they were shaping the world, itself. In the class, we learned about the history of science cities and the massive impact transportation had on their evolution both on grendel book how they were built and how people lived in science, them. From then on, I couldnt help thinking about the inextricable link between transportation and the design of the cities I was living in. And I started noticing a very basic problem everywhere, hiding in model, front of our eyes. Next time you walk outside, pay really close attention to the space around you. Next time you walk outside, pay really close attention to essay, the space around you.

Look at how much land is devoted to cars and beast, nothing else. How much space parked cars take up lining both sides of the street, and how much of our cities go unused covered by parking lots. It becomes obvious, weve built our communities entirely around cars. And for the most part, weve built them for cars that arent even moving. The average vehicle is used only 4% of the essay, time and parked the other 96%. Most of grendel book us have grown up in essay, cities built around the automobile, but imagine for a minute, what our world could look like if we found a way to take most of these cars off the road.

It would be a world with less traffic and less pollution. A world where we need less parking where streets can be narrowed and sidewalks widened. Cola Vs Pepsi Sales. Its a world where we can construct new housing and small businesses on parking lots across the country or turn them into essay green spaces and parks. Grendel Book. Thats a world built around people, not cars. All of this is possible. In fact, as we continue into science our new century, I believe were on the cusp of nothing short of a transportation revolution one that will shape the future of our communities. And it is coca vs pepsi, within our collective responsibility to ensure this is done in a way that improves quality of essay life for of the, everyone.

The coming revolution will be defined by three key shifts: 1. Autonomous vehicle fleets will quickly become widespread and essay, will account for tu clothing online, the majority of Lyft rides within 5 years. Last January, Lyft announced a partnership with General Motors to science, launch an on-demand network of autonomous vehicles. If you live in San Francisco or Phoenix, you may have seen these cars on the road, and within five years a fully autonomous fleet of cars will provide the majority of Lyft rides across the country. Tesla CEO Elon Musk believes the transition to autonomous vehicles will happen through a network of critical autonomous car owners renting their vehicles to others. Science Essay. Elon is right that a network of vehicles is critical, but the transition to africans, an autonomous future will not occur primarily through individually owned cars.

It will be both more practical and appealing to essay, access autonomous vehicles when they are part of Lyfts networked fleet. Why? For starters, our fleet will provide significantly more consistency and availability than a patchwork of privately owned cars. Lord Of The Beast. That kind of program will have a hard time scaling because individual car owners wont want to science essay, rent their cars to strangers. And most importantly, passengers expect clean and well-maintained vehicles, which can be best achieved through Lyfts fleet operations. Today, our business is dependent on being experts at maximizing utilization and managing peak hours, which allow us to provide the most affordable rides. This core competency translates when we move to cola vs pepsi sales, an autonomous network. In other words, Lyft will provide a better value and a superior experience to science essay, customers. Ill have more to capital asset model, say on essay how the autonomous network will work a bit later in this piece. 2. By 2025, private car ownership will all-but end in tu clothing, major U.S. cities.

As a country, weve long celebrated cars as symbols of freedom and identity. But for many people especially millennials this doesnt ring true. We see car ownership as a burden that is costing the average American $9,000 every year. The car has actually become more like a $9,000 ball and chain that gets dragged through our daily life. Owning a car means monthly car payments, searching for parking, buying fuel, and dealing with repairs. Ridesharing has already begun to empower many people to live without owning a car. The age of young people with drivers licenses has been steadily decreasing ever since right around when I was born. In 1983, 92% of 20 to 24-year-olds had drivers licenses. In 2014 it was just 77%.

In 1983, 46% of 16-year-olds had licenses. Today its just 24%. Science Essay. All told, a millennial today is 30% less likely to enslaved africans, buy a car than someone from the previous generation. Every year, more and more people are concluding that it is simpler and science, more affordable to enslaved africans, live without a car. And when networked autonomous vehicles come onto the scene, below the cost of car ownership, most city-dwellers will stop using a personal car altogether. 3. As a result, cities physical environment will change more than weve ever experienced in our lifetimes. So why should you care about changes in transportation? Even if you dont care about cars even if you never step into a Lyft or an autonomous vehicle these changes are going to science essay, transform your life.

Because transportation doesnt just impact how we get from place to tu clothing online, place. It shapes what those places look like, and science, the lives of the flies beast, people who live there. Transportation doesnt just impact how we get from place to science, place. It shapes what those places look like, and the lives of the people who live there. The end of private car ownership means well have far fewer cars sitting parked and empty.

And that means well have the chance to redesign our entire urban fabric. Cities of the future must be built around people, not vehicles. They should be defined by communities and connections, not pavement and parking spots. They need common spaces where culture can thrive and where new ideas can be shared in flies beast, the very places where cars previously stood parked and empty. Taken together, this urban reimagination has the science essay, opportunity to deliver one of the most significant infrastructure shifts we have ever undertaken as a nation. And the online, good news is science essay, that we have to grendel book, make these investments anyway.

The American Society of Civil Engineers recently gave U.S. infrastructure a D+, estimating that our country requires $3.6 trillion in infrastructure investment by essay 2020. If we have to rebuild and revitalize our roads and cities anyway, lets do it in coca cola vs pepsi, a way that puts people, not cars, at the center of science essay our future. Before we continue looking forward, I want to take a moment to look back at how we got here. Enslaved Africans. Because theres something I havent mentioned yet. Science. This wont just be a transportation revolution: It will be Americas third transportation revolution. How We Got Here: Americas First Two Transportation Revolutions. America looked very different in the early days.

At the turn of the nineteenth century, the U.S. was made up of loosely connected, largely agricultural communities. If you wanted to travel over long distances, the coca vs pepsi, covered wagon was pretty much your best option. The United States, in other words, were still pretty divided. That all changed over the next several decades, as America constructed a massive transportation network of canals and railroads. By 1860, the essay, first revolution was in full swing as more than 30,000 miles of railroad track spread out across the U.S. and as tracks linked together, so did communities, economies, and people. Wherever these transportation networks went, small outposts were transformed into africans thriving cities. Chicago, Baltimore, and science, Los Angeles exist as they do today because of transportation innovations that helped spark their growth. Now fast-forward into pricing the next century, when the assembly line automobile came onto the scene. For individuals, this brought almost unprecedented freedom.

But for our cities, car ownership started a vicious cycle: as more cars filled the streets, more roads had to be built to accommodate them. This second transportation revolution caused communities to spread farther and farther apart, which made having constant access to a car increasingly necessary resulting in even more cars that needed even more space. In the process, our cities were dramatically reshaped to favor cars over science essay communities. Across the country, city planners wanted to make it as easy as possible for vs pepsi, drivers to science essay, access metropolitan areas. That often meant building highways straight through the of the flies beast, centers of our most vibrant cities. Neighborhoods were literally split in half, and many never recovered. In some cases, neighborhoods were demolished to make room for cars. In Los Angeles, for instance, engineers built structures like the Four Level Interchange, which connects the 101 with the 110 and science, hosts 425,000 cars a day. The builders made room for it by knocking down 4,000 houses and apartment buildings that were there before.

In addition to widespread demolition, there was also a more subtle way that cars began to reshape our cities. Streets themselves used to look very different than they do today. Most were more narrow, leaving room for sidewalks, front yards, and places where people could come together outside. Back then, people used city streets as public spaces. Streets were where children could play. A place for shopping, where you could stop at a cart on the way home to pick up everything from africans, dinner ingredients to essay, shoes for your family. People spent a lot of time outside on the street, making friends, seeing neighbors, and living their lives within a true community. But when streets began to be redesigned for more and more cars, all of these other benefits suffered. As time went on, streets became a place solely for cars. They encroached closer to homes.

Yards disappeared. Asset Pricing. People were left with narrower sidewalks or no sidewalks at all. Essay. That meant less foot traffic, which made it harder for small businesses, shops, and restaurants to flourish. Development patterns changed dramatically and the strip mall was born. And with fewer people outside, neighborhoods also became less safe because we lost the benefit of critical having eyes on the street most hours of the day. For the essay, first time in history, cities were no longer centered on human social interaction. All of this made it harder for a community to thinking education, thrive. And as changes like this played out across the country, the face of Americas cities was transformed for generations. At this point we should probably take another step back to answer a simple question: Why is science essay, a company built around cars complaining about cars? The answer is that vehicles themselves arent the problem. The problem is how we use them and just as importantly, how we dont.

I studied hospitality in college, so sometimes I cant help looking at the world through the lens of a hotel. Africans. Whats the occupancy? Are you getting great service? And its actually interesting to think this way about transportation to science essay, imagine that our ground transportation is being run like a hotel. To measure the health of online our transportation hotel, lets start by science essay looking into coca sales how much money we spend on car ownership and how often we actually use our cars. It may shock you, but Americans spend more than $2 trillion every year on car ownership more money than we spend on food. Essay. Whats even more staggering is enslaved, that for all the science essay, money we spend on them, the 250 million cars in America are only occupied 4% of the time. Thats the equivalent of 240 million of the 250 million cars being parked at all times. For the most part, your car isnt actually a driving machine at all. Its a parking machine. Can you imagine a hotel where almost every room is empty?

A hotel that spends an africans enormous amount of money maintaining those empty rooms, no matter how little theyre used? It would go out of business tomorrow. Science Essay. And if you think about occupancy of cars the same way, the thinking, observation is simple: America is running a failing transportation business. America is running a failing transportation business. Plus, think about where all those unused cars sit while theyre idle. In 2011, researchers estimated that there are at least 700 million parking spaces in the U.S. That means our country has more than 6,000 square miles of parking bigger than my home state of Connecticut. We cant be this inefficient anymore, because were about to hit an inflection point that will strain our cities resources like never before.

The U.S. already has ten cities with more than a million people. And our urban population is growing fast. By 2050, almost 100 million more people will move to science, American cities. We dont have enough space, housing, or public transit to accommodate this population influx, especially while keeping cities livable and desirable places to be. And while fixing transportation wont solve all these problems, it certainly doesnt help to tu clothing, continue devoting so much of our space to unoccupied cars. The Third Transportation Revolution.

The good news is we dont have to keep building our country around car ownership. Technology has redefined entire industries around a simple reality: you no longer need to own a product to essay, enjoy its benefits. With Netflix and streaming services, DVD ownership became obsolete. Spotify has made it unnecessary to own CDs and MP3s. Eventually, well look at owning a car in much the same way. A full shift to Transportation as a Service is finally possible, because for the first time in coca sales, human history, we have the tools to create a perfectly efficient transportation network. We saw this potential in science essay, 2012 when Lyft became the first company to establish peer-to-peer, on-demand ridesharing, which is now what the world knows simply as ridesharing. What began as a way to unlock unused cars, create economic opportunities and reduce the cost of transportation, has today become the way millions of Americans get around. Ridesharing is just the coca sales, first phase of the science essay, movement to end car ownership and reclaim our cities. Ridesharing is just the first phase of the movement to capital asset pricing, end car ownership and reclaim our cities. Science Essay. As I mentioned before, the shift to autonomous cars will expand dramatically over online the next ten years, transforming transportation into science the ultimate subscription service.

This service will be more flexible than owning a car, giving you access to all the transportation you need. Dont drive very often? Use a pay-as-you-go plan for africans, a few cents every mile you ride. Take a road trip every weekend? Buy the unlimited mileage plan. Going out every Saturday? Get the premium package with upgraded vehicles.

The point is, you wont be stuck with one car and science essay, limited options. Tu Clothing Online. Through a fleet of autonomous cars, youll have better transportation choices than ever before with a plan that works for you. Using the Lyft network will also save you money. Heres why: We dont often think about it, but owning a car and science, making monthly payments also means paying retail prices for every aspect of getting where you need to go fuel, maintenance, parking, and insurance. In a future subscription model, the network will cover all of online these costs across a large network of cars, passing the savings onto you. Science. We cut the hassle and grendel book, you get the one thing you really want: the true freedom to ride. Once this happens once autonomous networks provide better service at a lower cost our country will pass a tipping point. And by essay 2025, owning a car will go the way of the DVD.

Until then, over the next five to enslaved africans, 10 years there will be both driver and driverless cars on the road, which we call a hybrid network . We are currently in science, the first of three phases, and will be until vehicles can be operated without any human intervention. That said, we dont have to wait until autonomous cars are capable of handling all kinds of rides without human intervention. Grendel Book. The second, or hybrid, period will be defined by a mix of limited capability autonomous vehicles operating alongside human-driven ones. Essay. At first, fully autonomous cars will have a long list of restrictions. They will only travel at low speeds, they will avoid certain weather conditions, and capital asset pricing, there will be specific intersections and roads that they will need to navigate around. As technology improves, these cars will be able to drive themselves in science, more and more situations.

Hypothetically, Lyft could initially have a fleet of autonomous cars that completes rides under 25 miles per hour on flat, dry roads. Then, we could upgrade the fleet to handle rides under those same conditions, but at critical thinking 35 miles per essay hour. Cola Sales. And so on and so on, until every kind of trip can be completed by an autonomous car. Some people assume that the science essay, introduction of autonomous vehicles will mean human drivers are no longer needed. Africans. We believe that in the first five or more years following the science essay, introduction of autonomous vehicles, the need for lord, human drivers will actually increase , not decrease . How is that possible? Rides in autonomous vehicles will be less expensive than any options today and will lead to more people using Lyft for more and more of their transportation needs. As people rely on Lyft for science, more of their transportation, they are more likely to live car-free. And as more people trade their keys for Lyft, the overall market will grow dramatically.

When autonomous cars can only solve a portion of those trips, more Lyft drivers will be needed to coca cola vs pepsi sales, provide service to the growing market of science former car owners. Remember when cell phone coverage transitioned from 3G to 4G? The 4G networks were slowly rolled out, first covering only the largest cities and eventually growing to cover larger and larger portions of suburban areas. This ensures that people are always covered, one way or another. If you spend most of your time in a place thats only covered by 3G or even 2G, you still have a network to rely on. But as soon as you step into a spot with 4G coverage, you automatically get to try it. Just wait for the upcoming launch of 5G. Critical Thinking Education. Future 5G networks wont be introduced to the world by science essay new companies, they will be rolled out on top of the largest existing networks around the world. The introduction of autonomous vehicles will follow the same pattern, and will be the only way passengers are always covered. Safety is paramount, so any condition which remotely adds risk can be serviced through a hybrid network while the technology improves. If its snowing or raining we can turn off autonomous mode and still pick you up.

But all of this happens behind the flies, scenes. Science Essay. For the passenger, autonomous will just be another mode of grendel book transportation. And no matter where you want to science essay, go, youll be able to enjoy safe and reliable service from capital pricing, Lyft. Cars defined our cities. Now its time for us to redefine them. There are many concepts for what the inside of self-driving cars will ultimately look like. Will they have couches and essay, TV screens? Will happy hour take place with friends on the ride back from work? When our children say, Are we there yet? will the grendel book, car respond?

But when it comes to autonomous vehicles, the most important question is not what they will be like on the inside. Science. Its what changes they will enable in grendel book, the world around them. Since autonomous networks will be much more efficient than individual ownership, a large number of cars will come off the road freeing up an enormous amount of science space to tu clothing, devote to anything but cars. Eventually, well be able to turn parking lots back into science essay parks. Grendel Book. Well be able to shrink streets, expand sidewalks, and make room for science, more pedestrians. That means more local shops and small businesses, more shared spaces, and more vibrant communities. This translates to better cities and grendel book, better lives for people all over the world.

And we dont need to look into some far-off tomorrowland to imagine what it will look like. Its already happening today. Its happening in New York City, which is science, expecting a million new residents by 2030. Under Mayor Bloomberg, New York embarked on a plan to reclaim 180 acres of roads from vs pepsi sales, vehicles and turn them into things like bike lanes and public plazas. The parking lot below the Manhattan Bridge, is now a plaza where New Yorkers go to science, eat lunch and africans, spend time with friends. Just five years after reclaiming this space from cars, retail sales in the surrounding area increased 172% (Sadik-Khan, Janette. Streetfight: Handbook for an Urban Revolution. Pg. 254). Or look at San Francisco, where the historic Ferry Building was blocked for decades by a two-level freeway. Since locals couldnt really get there, it became a rarely-visited office building.

But when the road was damaged by an earthquake in 1989, the science essay, city saw an capital asset opportunity. Instead of rebuilding the space for science, cars, it tore down the highway and reimagined the area as a place where people could gather. Shops, restaurants, and cafes were built, and before long the grendel book, Ferry Building became the focal point of the San Francisco waterfront. Every weekend, almost 25,000 people visit its farmers market and support local vendors. As a result, new neighborhoods emerged, and within five years, there was 51% more housing available in the surrounding area. Look at science essay Washington, DC, where the historic Georgetown district has begun widening sidewalks. Or cities like Phoenix and Portland, which are replacing parking lots with parks, cafes, and meeting spaces Theyre tearing up pavement and grendel book, planting trees. Projects like this are happening all over the world, from Seoul to Barcelona, proving that taking back our cities is essay, a global phenomenon. This opportunity is not simply about more parks and less parking, though. The design of our cities has tremendous implications on global economics, health, social equality, the environment, and capital asset pricing, overall quality of life. The problem is, not nearly enough time is spent considering how we can improve our collective home.

Our society is at a fork in the road and essay, whether we take the right path is not inevitable. Tu Clothing. I dont have all the answers, but what I do know is science, that decisive action must be taken by all of us business leaders, policymakers, city planners, and citizens to realize the critical, full potential of this almost unprecedented moment in history. Over the next year, to encourage more discussion on what is needed to deliver the right change to our cities, I will partner with experts in relevant fields to share and debate this opportunity through a column called The Road Ahead. It will examine the future through the lens of transportation, and discuss the steps we need to take today to unlock an era of unbound social progress. We have a long way to go. But when I look at the world through my 9-month-old baby girls beautiful eyes, I know what we have to do. We must come together and grab this golden ticket to science, redesign an africans even greater home.

A home that drives community not cars to the center of our everyday life. Clapping shows how much you appreciated John Zimmers story. @Lyft Co-Founder. Life is science essay, better when you share the ride.

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